oPhotometer®

nual
manual D30
gEN)

Register your instrument!

www.eppendorf.com/myeppendorf

Eppendorf BioPhotometer® D30

Operating manual
Copyright © 2014 Eppendorf AG, Hamburg. No part of this publication may be reproduced without the
prior permission of the copyright owner.

Trademarks

Eppendorf®, the Eppendorf logo, Eppendorf BioPhotometer®, Eppendorf μCuvette®, and UVette® are
registered trademarks of Eppendorf AG, Hamburg, Germany.

Cy® is a registered trademark of GE Healthcare UK Ltd., Buckinghamshire, UK.

Trademarks are not marked in all cases with ™ or ® in this manual.

This product is manufactured under license to issued U.S. Patent No. 6,122,052.

6133 900.053-02/052014
 Table of contents
Eppendorf BioPhotometer® D30 3
English (EN)

Table of contents
1 Operating instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.2.1 Danger symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.2.2 Danger levels. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.3 Symbols used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.4 Abbreviations used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

2 Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
2.1 Main illustration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
2.2 Delivery package . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
2.3 Features. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
2.3.1 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
2.3.2 Operation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
2.3.3 Result output . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
2.3.4 Device self test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

3 Safety. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
3.1 Intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
3.2 User profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
3.3 Warnings for intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
3.3.1 Personal injury . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
3.3.2 Damage to device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3.4 Information on product liability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
3.5 Safety instructions located on the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

4 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
4.1 Preparing installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
4.2 Selecting the location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
4.3 Connecting the device to the mains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
4.4 Connecting the printer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
4.4.1 Thermal printer DPU-S445 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
4.4.2 Thermal printer DPU-414 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
4.5 Connecting PC or USB stick for data export. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

5 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
5.1 Overview of operating controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
5.1.1 Entering text . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
5.2 Inserting the cuvette . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
5.3 Summary of the measuring procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
5.3.1 Preparing the measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
5.3.2 Measuring procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
5.3.3 Important measurement instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
 Table of contents
4 Eppendorf BioPhotometer® D30
English (EN)

6 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
6.1 Selecting a method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
6.2 Photometry method description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
6.2.1 Absorbance method group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
6.2.2 Routine method group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
6.2.3 Basic method group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
6.3 Method parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
6.4 Method procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
6.4.1 Check parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
6.4.2 Measure standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
6.4.3 Measure samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
6.4.4 Measure samples: Results displays. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
6.4.5 Process results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
6.4.6 Print & export . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
6.4.7 Finish the series of measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

7 Functions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
7.1 Functions of the User main group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
7.1.1 Results memory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
7.1.2 General method parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
7.1.3 Absorbance spectra library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
7.1.4 Device settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
7.1.5 Device calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
7.1.6 Info . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52

8 Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
8.1 Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
8.1.1 Cleaning the cuvette shaft cover . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
8.2 Disinfection/Decontamination. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
8.3 Checking the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
8.3.1 Checking the photometer unit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
8.3.2 Device self test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
8.4 Replacing fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
8.5 Decontamination before shipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59

9 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
9.1 General errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
9.2 Error messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
9.3 Result flags . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

10 Transport, storage and disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

10.1 Transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
10.2 Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
10.3 Disposal. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
 Table of contents
Eppendorf BioPhotometer® D30 5
English (EN)

11 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
11.1 Power supply. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
11.2 Ambient conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
11.3 Weight/dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
11.4 Photometric properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
11.5 Further technical parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
11.6 Application parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71

12 Evaluation procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
12.1 Absorbance values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
12.1.1 Blank . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
12.1.2 Background correction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
12.1.3 Cuvette correction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
12.2 Evaluation with factor or standard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
12.3 Evaluation with standard curve/line . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
12.4 Dilution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
12.5 Special evaluation procedures for nucleic acids and protein UV . . . . . . . . . . . . . . . . . . . . . . . . 77
12.5.1 Ratios A260/A280 and A260/A230.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
12.5.2 Conversion to molar concentrations and nucleic acid quantities . . . . . . . . . . . . . . . . . 77
12.5.3 Calculating the factor for protein in "General Method Parameter" . . . . . . . . . . . . . . . 79

13 Ordering information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81

Certificates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
 Table of contents
6 Eppendorf BioPhotometer® D30
English (EN)
 Operating instructions
Eppendorf BioPhotometer® D30 7
English (EN)

1 Operating instructions
1.1 Using this manual

 Read this operating manual completely before using the device for the first time. Also observe the
instructions for use of the accessories.
 This operating manual is part of the product. Thus, it must always be easily accessible.
 Enclose this operating manual when transferring the device to third parties.
 You will find the current version of the operating manual for all available languages on our webpage
under www.eppendorf.com.

1.2 Danger symbols and danger levels

The safety instructions of this operating manual indicate the following danger symbols and danger levels:

1.2.1 Danger symbols

Electric shock Explosion

Toxic substances Hazard point

Material damage

1.2.2 Danger levels

DANGER Will lead to severe injuries or death.

WARNING May lead to severe injuries or death.
CAUTION May lead to light to moderate injuries.
NOTICE May lead to material damage.

1.3 Symbols used

Depiction Meaning
1. Actions in the specified order
2.
 Actions without a specified order
• List
Additional information
 Operating instructions
8 Eppendorf BioPhotometer® D30
English (EN)

1.4 Abbreviations used

A
Absorbance

DNA
Deoxyribonucleic acid

dsDNA
Double-stranded DNA

M
mol/L (molar)

OD600
Optical density at a wavelength of 600 nm

RNA
Ribonucleic acid

ssDNA
Single-stranded DNA

UV
Ultraviolet radiation

Vis
Visible light

CV
Coefficient of variation (standard deviation/average value) in percent
 Product description
Eppendorf BioPhotometer® D30 9
English (EN)

2 Product description
2.1 Main illustration
Abb. 2-1: BioPhotometer D30: Front view and rear view

1 2 3 3

ce
absorban

absorban
ce
height
8.5 m m

1 2
abc 3
def
meth od
ghi
4 5
jkl
mno
6 exit
func tion
7
pq rs 8
tuv
wxyz
9 0 dele te
µ %

ente r standard
blan k
sample

10 9 8 7 6 5 4

Fig. 2-1: BioPhotometer D30: Front view and rear view

1 Display 6 Fuse holder

2 Cuvette shaft 7 Mains/power connection
3 Cuvette shaft cover 8 USB connection for PC
4 USB connection for USB stick and printer 9 Connection for RS-232 printer
5 Mains/power switch 10 Operating controls

The name plate is located at the bottom left on the underside of the device.

2.2 Delivery package

Quantity Description
1 BioPhotometer D30
1 Power cord
4 4 UVettes
Original Eppendorf plastic cuvette, individually packaged, PCR clean, protein-free
1 Operating manual, in multiple languages
 Product description
10 Eppendorf BioPhotometer® D30
English (EN)

2.3 Features

The BioPhotometer D30 is a UV VIS photometer for measuring liquids in cuvettes. As the measuring data
are collected at fixed wavelengths, the device especially is suited for routine applications in the
biomolecular, biotechnological, biochemical and cytological areas in research and development.

2.3.1 Methods

Preprogrammed methods and method templates

• Concentration determination of nucleic acids and proteins
• Determining the bacterial density by turbidity measurements: Method OD 600
• Method templates for different measurement and evaluation procedures:
– Quick absorbance measurements
– Evaluations with factor, standard and standard curve
• It is possible to create individual methods on the basis of the preprogrammed methods and templates.
• Quick measurement of absorbances without any further evaluation: Absorbance method group.

2.3.2 Operation

The preprogrammed methods and templates are combined into clearly arranged groups from which the
desired method can be quickly selected. After calling up the method, you are guided through the
measuring procedure in clear steps. If required, a help box in the display will provide you with hints. The 3
measuring keys (standard, blank, sample) enable you to start the measurement quickly and directly.

2.3.3 Result output

The BioPhotometer D30 emits the results on the display as well as via a printer which is available at
Eppendorf. With a USB connection, you can transfer result data from the device to a USB stick, a printer or
directly to a PC.

2.3.4 Device self test

Directly after switching on, the device checks the functioning of the photometer unit by itself. Access the
Device calibration function for a more comprehensive test (see Device self test on p. 58).
 Safety
Eppendorf BioPhotometer® D30 11
English (EN)

3 Safety
3.1 Intended use

The BioPhotometer D30 is to be used in molecular biology, biochemistry and cell biology research
laboratories. The BioPhotometer D30 exclusively is determined for use in the interior of buildings. All
country-specific safety requirements for operating electrical equipment in the laboratory must be observed.

The BioPhotometer D30 is used for photometric concentration determination of biomolecules in liquids as
well as for turbidity measurements of microbiological cultures in routine laboratories.

Only use Eppendorf accessories or accessories recommended by Eppendorf.

3.2 User profile

The device and accessories may only be operated by trained and skilled personnel.

Before using the device, read the operating manual carefully and familiarize yourself with the device's
mode of operation.

3.3 Warnings for intended use

3.3.1 Personal injury

DANGER! Electric shock as a result of penetration of liquid.

 Switch off the device and disconnect the power plug before starting cleaning or
disinfection work.
 Do not allow any liquids to penetrate the inside of the housing.
 Do not spray clean/spray disinfect the housing.
 Only plug the device back in if it is completely dry, both inside and outside.

DANGER! Risk of explosion.

 Do not operate the device in areas where work is completed with explosive substances.
 Do not use this device to process any explosive or highly reactive substances.
 Do not use this device for processing any substances which could generate an explosive
atmosphere.

WARNING! Electric shock due to damage to device or mains cable.

 Only switch on the device if the device and mains cable are undamaged.
 Only use devices that have been properly installed or repaired.
 In case of danger, disconnect the device from the mains supply by pulling the power plug
from the device or the mains socket or, by using the isolating device intended for this
purpose (e.g., emergency stop switch in the laboratory).
 Safety
12 Eppendorf BioPhotometer® D30
English (EN)

WARNING! Damage due to UV radiation.

Microliter cuvettes, e.g., Hellma® TrayCell (or microliter cuvettes with a similar design) divert
the radiation from the light source within the cuvette so the radiation can escape upward
when the lid is not closed.

 Before starting a measurement, ensure that the lid on the microliter cuvette is not open.

WARNING! Damage to health from toxic, radioactive or aggressive chemicals as well as

infectious liquids and pathogenic germs.

 Observe the national regulations for handling these substances, the biological security
level of your laboratory, the material safety data sheets and the manufacturer's application
notes.
 Wear personal protective equipment.
 For full instructions regarding the handling of germs or biological material of risk group II
or higher, please refer to the "Laboratory Biosafety Manual" (Source: World Health
Organization, current edition of the Laboratory Biosafety Manual).

WARNING! Damage to health due to contaminated device and accessories.

 Decontaminate the device and the accessories before storage and shipping.

CAUTION! Poor safety due to incorrect accessories and spare parts.

The use of accessories and spare parts other than those recommended by Eppendorf may
impair the safety, functioning and precision of the device. Eppendorf cannot be held liable or
accept any liability for damage resulting from the use of incorrect or non-recommended
accessories and spare parts, or from the improper use of such equipment.

 Only use accessories and original spare parts recommended by Eppendorf.

3.3.2 Damage to device

NOTICE! Damage from the use of aggressive chemicals.

 Do not use any aggressive chemicals on the device or its accessories, such as strong and
weak bases, strong acids, acetone, formaldehyde, halogenated hydrocarbons or phenol.
 If the device has been contaminated by aggressive chemicals, immediately clean it by
means of a mild cleaning agent.

NOTICE! Damage to the device from fumigating with aggressive chemicals.

 Do not use fumigation to disinfect the device.

 Safety
Eppendorf BioPhotometer® D30 13
English (EN)

NOTICE! Corrosion from aggressive cleaning agents and disinfectants.

 Do not use corrosive cleaning agents, aggressive solvents or abrasive polishes.

 Do not incubate the accessories in aggressive cleaning agents or disinfectants for a longer
period of time.

NOTICE! Damage to electronic components due to condensation.

Condensate can form in the device after it has been moved from a cool environment to a
warmer environment.

 After installing the device, wait at least for 3 h. Only then connect the device to the mains.

NOTICE! Function impairment due to mechanical damage.

 After mechanical damage to the device, ensure that the measuring and evaluation
functions of the device are operating correctly by completing an inspection.

NOTICE! Damage from overheating.

 Do not install the device near to any heat sources (e.g., heating, drying cabinet).
 Do not expose the device to direct sunlight.
 Ensure unobstructed air circulation. Keep free a clearance of at least 5 cm around all
ventilation grilles.

NOTICE! Material damage from incorrect use.

 Only use the product for its intended purpose as described in the operating manual.
 Ensure adequate material resistance when using chemical substances.
 In case of doubt, contact the manufacturer of this product.

NOTICE! Damage as a result of incorrect packing.

Eppendorf AG is not liable for damage caused by improper packing.

 The device may only be stored and transported in its original packaging.

NOTICE! Damage due to improper cleaning of the cuvette shaft.

 Only clean the cuvette shaft using a moist cotton swab .

 Do not allow any liquid to enter the cuvette shaft.
 Do not reach with your fingers into the cuvette shaft.
 Safety
14 Eppendorf BioPhotometer® D30
English (EN)

3.4 Information on product liability

In the following cases, the designated protection of the device may be compromised. Liability for any
resulting property damage or personal injury is then transferred to the operator:

• The device is not used in accordance with the operating manual.

• The device is used outside of its intended use.
• The device is used with accessories or consumables which are not recommended by Eppendorf.
• The device is maintained or repaired by people not authorized by Eppendorf.
• The user makes unauthorized changes to the device.

3.5 Safety instructions located on the device

Depiction Meaning Location
Hazard point Rear side of the device

 Follow the operating manual.

The device needs to be readjusted Bottom of the device

Gerät nach dem Öffnen after it has been opened.
justieren!

Adjust device after

opening!  Do not open the device.
 Installation
Eppendorf BioPhotometer® D30 15
English (EN)

4 Installation
4.1 Preparing installation

 Keep the transport carton and the packing material for subsequent safe transport or storage.
 Check the completeness of the delivery using the information in the delivery package (see Delivery
package on p. 9).
 Check all parts for any transport damage.

4.2 Selecting the location

Select the location for the BioPhotometer D30 according to the following criteria:

• 2 grounded sockets for the BioPhotometer D30 and for the printer.
• Solid laboratory bench with horizontal work surface
Space requirement of the device: 50 cm (with printer: 75 cm) width, 50 cm depth.
• Temperature: 15°C to 35°C.
• Avoid temperature fluctuations (e.g, caused by open windows).
• Avoid direct sunlight.
• Humidity: 25% to 70% relative humidity.

Ensure that no objects (e.g., loose sheets, notebooks) that could impede the flow of air are
positioned under the device.

4.3 Connecting the device to the mains

1. Place the BioPhotometer D30 on a suitable work surface.
2. Verify that the mains/power supply voltage and mains/power frequency match the information on the
name plate.
3. Connect the device to the mains/power line and switch it on with the power switch.
4. Remove the protective film from the display.
 Installation
16 Eppendorf BioPhotometer® D30
English (EN)

4.4 Connecting the printer

4.4.1 Thermal printer DPU-S445

Prerequisites
Software version 3.4.4.0 or higher is installed on the device.

Connect the thermal printer DPU-S445 to the USB port for printers.

1. Connect the printer cable with the USB port for printers 4 (see Main illustration on p. 9).
2. Connect the printer cable with the printer.
3. Connect the printer to the mains/power line using the supplied mains/power adaptor and mains/power
cord (printer accessory) and switch it on.
For information on the printer, refer to the operating manual of the printer.

4.4.2 Thermal printer DPU-414

Connect the thermal printer DPU-414 to the serial printer connection.

1. Connect the printer cable to the serial printer connection 9 and tighten the locking screws.(see Main
illustration on p. 9).
2. Connect the printer cable to the printer and tighten the locking screws as well.
3. Connect the printer to the mains/power line using the supplied mains/power adaptor and mains/power
cord (printer accessory) and switch it on.
Information about modifying printer settings can be found in the operating manual for the printer.

The DIP switches are preset for the BioPhotometer D30 according to the following table.

Tab. 4-1: Setting the DIP SW for the thermal printer

DIP SW-1 Meaning
1 (OFF) Input = Serial
2 (ON) Printing Speed = High
3 (ON) Auto Loading = ON
4 (OFF) Auto LF = OFF
5 (ON) Setting Command = Enable
6 (OFF) Printing
7 (ON) Density
8 (ON) = 100%

DIP SW-2 Meaning

1 (ON) Printing Columns = 40
2 (ON) User Font Back-up = ON
3 (ON) Character Select = Normal
 Installation
Eppendorf BioPhotometer® D30 17
English (EN)

DIP SW-2 Meaning

4 (ON) Zero = Normal
5 (ON) International
6 (ON) Character
7 (ON) Set
8 (OFF) = U.S.

DIP SW-3 Meaning

1 (ON) Data Length = 8 bits
2 (ON) Parity Setting = NO
3 (ON) Parity Condition = Odd
4 (OFF) Busy Control = XON/XOFF
5 (OFF) Baud
6 (ON) Rate
7 (ON) Select
8 (ON) = 9600 bps

4.5 Connecting PC or USB stick for data export

You can connect a FAT 32-formatted USB stick to the USB port 4 (see Main illustration on p. 9).

Alternatively, you can connect the device for the data export directly to a PC by using a USB cable:

Prerequisites
• PC with Windows, version XP, SP2 or higher version.
• USB cable with a type A and type B plug each.
 Connect the device to the PC by using the USB cable on the USB port 8 (see Main illustration on p. 9).
• You do not need any special PC software for the data transmission: the transferred data
packets are recognized by the PC like a USB stick as a removable medium. For viewing the
data, you only need to open the registered data packet.
• The transmission of data to the USB stick or to the PC is started after completing the series
of measurement in the print & export (see Print & export on p. 43) method step.
 Installation
18 Eppendorf BioPhotometer® D30
English (EN)
 Operation
Eppendorf BioPhotometer® D30 19
English (EN)

5 Operation
5.1 Overview of operating controls
Abb. 5-1: Control panel of the BioPhotometer D30

Fig. 5-1: Control panel of the BioPhotometer D30

Key: Function

Keypad: Enter digits and text.

Keys 1 to 9 as well as 0: When entering text, next to numbers you also can
enter letters and special characters by pressing the key several times.
Alternatively, you can switch to a displayed keyboard with the [Keyboard] key.

Outside of entry fields: Call up method selection.

Outside of entry fields: Call up function selection.

Softkey: Select functions.

The key assignment changes along with the software dialog. The current
function is displayed directly above the key on the display.

Move the cursor to the left, right, up, down.

• Navigation between input fields.
• and keys inside an entry field: Navigate within the character string.
• and keys in a result display: Navigate between the sample results of
the series of measurement.
• and keys within a graph: Navigate on the x-axis of the graph, e.g. for
displaying the wavelength-dependent absorbance values in a scan.
 Operation
20 Eppendorf BioPhotometer® D30
English (EN)

Key: Function

Exit the current selection for the next higher level.

Delete entry. Within a sequence of signs, the sign on the left of the cursor is
deleted
• Call up selected method or function.
• Open the selection list.
• Confirm entry or selection.

Start standard measurement.

Start blank measurement.

Start sample measurement.

 Operation
Eppendorf BioPhotometer® D30 21
English (EN)

5.1.1 Entering text

You can enter texts when assigning method names and result units. Restriction: Only digits, letters and the
underscore "_" are allowed for method names.

Entry via keyboard:

Use the and cursor keys to navigate within the
entry field and to change single positions in the
name.
Softkeys:
• [Keyboard]: Display keyboard.
• [abc]: Change between upper and lower case
letters when making entries with the keypad.
• [Save]: Save entered text.
• [Cancel]: Cancel text input.

Entry via the displayed keyboard:

Use the cursor keys to select the displayed signs and
respectively confirm your selection with the enter
key. As for a PC key pad, you can use the "Shift"
resp. the "Caps Lock" key for changing the
capitalization for the next entry or for all following
entries.
Softkeys:
• [Numbers]: Switch to entry using the keyboard.
• [Save]: Save entered text.
• [Cancel]: Cancel text input.
 Operation
22 Eppendorf BioPhotometer® D30
English (EN)

5.2 Inserting the cuvette

Standard rectangular glass or plastic cuvettes can be inserted in the cuvette shaft:

• External dimensions: 12.5 mm × 12.5 mm

• Height of light path: 8.5 mm higher than cuvette base
• Total height: min. 36 mm

The cuvettes must be optically transparent for the respective measuring wavelength. For measurements in
the UV range, Eppendorf offers the plastic cuvette UVette which is transparent for wavelengths of 220 nm
and higher and therefore also is suitable for measuring nucleic acids.

Eppendorf Hellma ® TrayCell * UVette ® Ultra-micro Semi-micro Macro

Cuvettes μCuvette G1.0
Basic area 12.5 mm × 12.5 mm
Min. overall height 36 mm

Min. filling level 10 mm

Light path 8.5 mm
Max. height of base 7 mm
0 mm
Min. volume Photometry See manufacturer See manufacturer 50 μL 70 μL 400 μL 1000 μL
information information
* or similar microliter cuvette

Prerequisites
• The cuvette is free from contamination by dust or fingerprints and free from scratches.
• The cuvette shaft is free from particles, dust and liquid.
• The measuring volume in the cuvette is sufficient. Ensure that the minimum measuring volume has
been reached.
• The measuring solution is free from particles and bubbles.
• The cuvette temperature and temperature control of the cuvette are above the temperature of the dew
point that applies for the ambient conditions (humidity and temperature).

The direction of the light path is marked with an arrow on the housing.

1. Position the cuvette so that the optical window of the cuvette is pointing towards the direction of the
light path.
2. When inserting the cuvette, press it completely to the bottom against the slight resistance.
 Operation
Eppendorf BioPhotometer® D30 23
English (EN)

5.3 Summary of the measuring procedure

5.3.1 Preparing the measurement

1. Switch on the device and, if required, the printer.

The device performs a self test (taking approx. 1 minute) and displays the method selection.
2. Make ready the cuvettes for the measurements (see Inserting the cuvette on p. 22).
3. Prepare the measuring solutions for measuring the blank values, if required, also the standards and the
samples.
4. Open the cover of the cuvette shaft. The cover can remain open during the measurements.
You should not use any measuring solution for standards and samples with a lower
absorbance than 0.02 to 0.03 A (e.g. dsDNA concentration between 1.0 and 1.5 μg/mL). The
detection limit of the device may be significantly lower, nevertheless, the impact of
disturbances from the measuring solutions (particles, bubbles, turbidity) on the reliability of
the result is very high for these low absorbance values.

5.3.2 Measuring procedure

5.3.2.1 Selecting a method

 Use the cursor keys to select the desired

method and call up the method with the enter
key.
For an overview and a detailed description of the
methods, refer to the next chapter (see Methods on
p. 29).

Wizard: The wizard at the top of the display will

take you through the method procedure
step-by-step.
Help box: You will receive help texts in the lower
right of the display during each step of the
procedure.
Softkeys: The [< Back] and [Next >] softkeys allow
you to move between method steps in the wizard.
 Operation
24 Eppendorf BioPhotometer® D30
English (EN)

5.3.2.2 Checking parameters

 Check the parameter setting. The [Page dn] and

[Page up] softkeys allow you to call up the
parameter list pages. You can modify and save
parameters using [Edit].

5.3.2.3 Measuring the blank and standards

For evaluations without standards (e.g. DNA measurements), this method step is omitted.

1. Start by measuring a blank (blank key).

2. Then measure all standards one by one
(standard key).
The display always marks the standard that is to be
measured next. Use the [Graph] resp. [Table]
softkey to change the result view.

 Press [Next] to accept the evaluation calculated

from the standard results.
 Operation
Eppendorf BioPhotometer® D30 25
English (EN)

5.3.2.4 Measuring samples

 The sample key is used for measuring your

samples consecutively.
Blank results will remain saved for the duration of
one series of measurement. However, a new blank
measurement always is possible. (The adjacent
figure shows a measuring procedure with
evaluation via the standard curve and, in addition
to the sample result, the graph of the standard
evaluation.)

5.3.2.5 Finalizing the method

1. Press [Finish], to complete the measuring series

and return to the method selection.
2. After all measurements have been completed,
switch off the device and close the cuvette shaft
cover to protect the cuvette shaft from
contamination.

5.3.2.6 Optional: process results

For the methods of the Nucleic acids method

group, you can postprocess the results in the
process results method step.

 Use the and cursor keys for selecting

systematically any results of the series of
measurement for postprocessing.
More Calc. softkey: Convert the concentration
results to molar concentrations or to nucleic acid
quantities (unit mass or mol).
 Operation
26 Eppendorf BioPhotometer® D30
English (EN)

5.3.2.7 Printing and exporting

1. Compose data packets for all samples or for

selected samples.
2. Print the data, save them to a USB stick or
transfer them to a PC via a USB cable.
 Operation
Eppendorf BioPhotometer® D30 27
English (EN)

5.3.3 Important measurement instructions

Check for each measurement:

• For plastic cuvettes: How many consecutive measurements can be reliably carried out in
the cuvette?
• Measure the cuvette blank value before the sample or standard measurements in order to
compensate the cuvette blank value in addition to the reagent blank.
• Blank results remain saved for one measuring series, but a new blank result measurement
can be performed at any time, even between sample measurements.
• The displayed absorbance values always correspond to the directly measured values. The
dilution or cuvette factor as well as background absorbances only will be incorporated for
the following result calculation (see Absorbance values on p. 73).
• The measuring result is typically displayed 2 to 3 seconds after a measurement has been
started. If (for high absorbance values) only a low quantity of light reaches the receiving
part, the measuring time automatically can be extended to 9 seconds in order to increase
the precision of the measurement.
• Observe that the measured absorbance values do not exceed the upper limit of the
photometric measuring range. In this case, reject the measuring result. The upper limit of
the photometric measuring range does not only depend on the wavelength (see
Photometric properties on p. 70) but also on the cuvette blank. Ultra-micro cuvettes with a
small diaphragm, such as TrayCell (Hellma), may have a cuvette blank of approx. A = 1.
The available photometric measuring range is reduced by this amount. You can estimate
the cuvette blank by measuring the cuvette filled with demineralized water as a sample in
comparison with the empty cuvette shaft as a blank. The cuvette blank of the Eppendorf
μCuvette G1.0 is negligible (approximately A = 0).
• After the measurement, remove the measuring solution completely before filling in the
next measuring solution in order to minimize carry-over. If a carry-over from one sample to
the next sample can be expected due to a high concentration difference, rinse the cuvette
between the measurements.
• If the temperature between the lamp and the ambience differs, photometric drift may
occur. Therefore a device from a colder ambience first has to be adjusted to the ambient
temperature.
Avoid quick changes of temperature. Carry out a new blank measurement for a long series
of measurements or measurements over a long period of time.
 Operation
28 Eppendorf BioPhotometer® D30
English (EN)
 Methods
Eppendorf BioPhotometer® D30 29
English (EN)

6 Methods
6.1 Selecting a method

Methods and method templates are delivered preprogrammed. The methods are organized in main groups
and subgroups.

Write-protected methods The most important methods in molecular biology.

Parameters can be modified, but the modified parameters
must be saved under a new method name.
Non-write-protected methods You can change parameters any number of times and start
the measurement right after saving.
New methods ("templates") Each method group contains a template which is
preprogrammed with complete parameter sets to facilitate
the programming of new methods. The parameters can be
changed and saved under new names any number of times.

To call a method, first use the cursor keys to select the main group, subgroup and the method. Confirm
each with enter.

Tab. 6-1: Photometric methods

Absorbance Methods for fast, simple absorbance measurements without additional
evaluations
Routine Frequently used molecular biology methods. The methods are
preprogrammed. However, the parameters can be modified if saved under
a new name.
Basic Methods for the evaluation of absorbance measurements with factor,
standard or standard curve/line.
Favorites In Favorites, you can set up your own folders using <New folder>, and
copy your frequently used methods to this folder in order to quickly
access them when needed.
 Methods
30 Eppendorf BioPhotometer® D30
English (EN)

You can create new methods in all folders using <New Method>.

In Favorites, you can create your own folders (e.g., to allocate folders to specific people), and rename and
delete the folders.

Tab. 6-2: Softkeys in method selection

[Cut] and [Paste] Cut and paste methods.
[Copy] and [Paste] Copy and paste methods.
[Delete] Delete methods.
[Rename] Rename methods.

Copied or cut methods can be added to a different folder under Favorites, or added to the original folder
under a new name. Use the cursor keys to navigate to the Methods column of the desired folder and press
[paste] for adding the method.

6.2 Photometry method description

The preprogrammed methods and method templates are described in this section.

6.2.1 Absorbance method group

Single λ
• Absorbance measurement on a wavelength.
• No subsequent evaluation.

6.2.2 Routine method group

The methods for the Routine group are preprogrammed as fixed methods. Therefore, a new method name
is required after the method parameters in the fixed preprogrammed methods have been modified.

Nucleic acids
• Determination of the concentration of nucleic acids through measurement at 260 nm and evaluation via
factor.
• Various nucleic acid methods, such as dsDNA or RNA, are preprogrammed. The parameters vary
according to the factor.
• Preprogrammed method for microliter cuvettes: Measuring DNA in sample volumes within the
microliter range with 1 mm light path (with microliter cuvettes as Eppendorf μCuvette G1.0 or Hellma®
TrayCell).
• Additional information on the purity of the measured nucleic acid: Ratios A260/A280, ratios A260/A230,
restricted absorbance wavelength spectrum of nucleic acid (with 3 nm distances), absorbance of the
background wavelength (preset: 320 nm; the absorbance of the pure nucleic acid should be close to
zero here).
• Partial turbidity correction can be performed via the Background parameter.
• Concentrations can be converted to molar concentrations and (after the sample volume has been
entered) to nucleic acid quantities (process results method step).
 Methods
Eppendorf BioPhotometer® D30 31
English (EN)

Proteins direct UV
• Determination of the concentration of proteins via measurement at 280 nm and factor or standard
evaluation.
• Preprogrammed methods for direct absorbance output as a result (Protein A 280) and for evaluation via
albumin-specific absorbance coefficients (Albumin A 280).
• Preprogrammed method for microliter cuvettes: Measuring protein in sample volumes in the microliter
range with 1 mm light path (with microliter cuvettes as Eppendorf μCuvette G1.0 or Hellma® TrayCell).
• Additional information on the purity of the measured protein: Absorbance of the background
wavelength (preset: 320 nm; the absorbance of the pure protein should be close to zero here).
• Partial turbidity correction can be performed via the Background parameter.
• When programming the methods, the corresponding factor is imported through the simple selection of
the protein from a predefined list. The factors are separately defined in the functions of the Gen.
method param. group. Various proteins are preprogrammed in Gen. method param.; additional
proteins can be added.

Proteins (with reagent)

• Concentration determination of proteins via measurement according to color reactions and evaluation
using standards or factors (typical: evaluation with standard curve).
• The Bradford, Bradford micro, Lowry, Lowry micro, BCA and BCA micro methods are already
preprogrammed. According to the reagent manufacturer, the "Curve fit" (standard curve type) must be
changed as necessary.

Bacterial density
• Turbidity measurement to determine the bacteria density.
• Measurement at 600 nm is already preprogrammed.

6.2.3 Basic method group

Factor, standard
• Measurement on a wavelength and factor or standard evaluation.
• Methods for factor and standard evaluation are preprogrammed.

Calibration curve
• Measurement on a wavelength and subsequent evaluation with a series of 2 to 12 standards.
• You can select between different evaluation procedures ("Curve fit") as linear regression, non-linear
regression.
• Graphical and tabular display of the standard results.
• The last saved standard evaluation can be used.
• A method for standard curve evaluation is preprogrammed.
 Methods
32 Eppendorf BioPhotometer® D30
English (EN)

6.3 Method parameters

This chapter illustrates the parameters for programming the methods. The order of the parameters in the
device display may slightly differ from the order in the table in order to display the parameters more clearly.
The table displays all parameters available for the various methods. Only a small portion of these
parameters are required for the corresponding method and will be shown in the display.

Parameter Entry Explanation

Cuvette Selection: Optical path length of the cuvette. The device always
10 | 5 | 2 | 1 | 0.5 | 0.2 | automatically converts absorbance values to the 10 mm path
0.1 mm length of a standard cuvette (see Absorbance values on p. 73).
Therefore, there is no need to change factors such as "50" for
the calculation of dsDNA concentrations when modifying the
Cuvette parameter.
Wavelength Selection: Measurement wavelength: The concentration is calculated
230 | 260 | 280 | 320 | based on the absorbance measured with this wavelength.
340 | 405 | 490 | 562 | For some method groups (e.g., Nucleic acids and Proteins
595 | 600 nm direct UV), the wavelengths are preprogrammed.
Unit Selection: Unit for the concentration result.
mg/mL | μg/mL | ng/ In the preprogrammed methods of the Routine group, the
mL | pg/mL | μg/μL | selection is restricted to units that are useful for these methods.
mg/dL | μmol/mL |
nmol/mL | pmol/mL |
pmol/μL | U | U/mL |
U/L | % | Abs | A/min
In addition, further
units are freely
programmable in the
General Method
Parameters/Units
function. Max. 7 digits.
Calculation Selection: Evaluation procedure for the calculation of the sample
factor | standard concentration from the measured absorbance.
Factor Value input: Factor for converting absorbance values into the concentration.
Factor. For the Factor method group, you also can enter negative
Limit: max. of 6 digits factors.
including decimal
point.
Protein Selection: Only for the Proteins direct UV method group.
List of protein types When selecting the protein, the corresponding Factor
which are stored in the parameter programmed in the General Method Parameters/
General Method Proteins function also will be imported from that function.
Parameters/Proteins
function.
 Methods
Eppendorf BioPhotometer® D30 33
English (EN)

Parameter Entry Explanation

Standards Value input: Number of different standard concentrations for the evaluation
Number of standards. with standards.
Range: 1 to 12. For some methods the range for the number of standards is
restricted to a smaller range than 1 to 12.
Replicates Value input: Number of repeated measurements for the various standard
Number of replicates concentrations.
per standard.
Range: 1 to 3.
Std. conc. Value input: Based on the number of standards, this parameter is available
Concentration values for all standards (e.g.,: std. conc. 1, std. conc. 2, ...).
of the standards.
Limit: max. of 6 digits
including decimal
point.
Decimal places Value input: Number of decimal points for the calculated concentration
Number of decimal result.
points for the result.
Range: 0 to 3.
Show scan Selection: Only for the Nucleic acids and Proteins direct UV method
on | off groups: Display of a restricted scan
(absorbance-wavelength-graph with distances of 3 nm) in
addition to the result of the sample measurement.
A260/A280 Selection: Only for nucleic acids.
on | off Display of the A260/A280 ratio in addition to the result of the
sample measurement.
A260/A230 Selection: Only for nucleic acids.
on | off Display of the A260/A230 ratio in addition to the result of the
sample measurement.
Background Selection: Only for the Nucleic acids and Proteins direct UV method
on | off groups:
Prior to the calculation of the results of a sample the
absorbance of a background wavelength, during which the
analyte to be measured should exhibit the absorbance value
zero, is subtracted from the absorbance of the measuring
wavelength. Frequent application: Partial correction of
turbidity for measurement of nucleic acids (background
wavelength in this case: 320 nm or 340 nm).
Wavelength Selection: 320 | Wavelength at which the background is to be measured. The
340 nm analyte to be measured should have the absorbance value zero
in pure form here.
 Methods
34 Eppendorf BioPhotometer® D30
English (EN)

Parameter Entry Explanation

Autoprint Selection: Printing a measuring result immediately following
on | off measurement with the thermal printer.
Only the main result data will be printed. To output detailed
data, the required data packets can be compiled and printed in
the print & export method step at the end of a measuring
series.

6.4 Method procedure

Wizard: the wizard at the top of the display will take
you through the method procedure. The currently
active method step is highlighted.

A method procedure is composed of a maximum of 5 steps. The currently active step is highlighted visually.
After the last step, print & export, of a measuring series, the start of a new measuring series is offered as a
next step. It once again starts with the sample measurement.

Method step Explanation

Check parameters Check method parameters. Carry out changes if required.
Measure standards Only for methods with standard evaluation:
Measure and evaluate standards. Alternatively, the last saved standard
evaluation can be used.
Measure samples Measuring samples
Process results Only for methods of the Nucleic acids group: postprocess results (conversion
of the concentration results).
Print & export Assemble data packets for printing or exporting the data.

Use the [Next >] and [< Back] softkeys to navigate between method steps. With [Abort] and [Finish] you can
cancel or finish the measuring procedure. The name of this softkey changes from [Abort] to [Finish] after
the first sample measurement.
 Methods
Eppendorf BioPhotometer® D30 35
English (EN)

6.4.1 Check parameters

Softkeys
• [Page dn] and [Page up]: Change between 1 to 3
parameter list pages.
• [Edit]: Switch to the parameter edit mode.

Editing mode for parameters:

Modified parameters are marked with a red star until
the modification has been saved.

Softkeys
• [Save] and [Save as]: Save changes. When using
[Save as] you have to rename the method. This is
always the case when modifying the methods
preprogrammed by Eppendorf in the Routine
group.
• [Cancel]: Exit the edit mode without saving
changes.
Saving the method under a new name:
You can save the method in the same folder from
which you called up the method or in any folder in
the Favorites method group.
The name (maximum 20 letters) can be entered
using a displayed keyboard ([Keyboard] softkey) or
directly using the keyboard (see Entering text on
p. 21).
After saving you will return to the check parameters
display.
 Methods
36 Eppendorf BioPhotometer® D30
English (EN)

6.4.2 Measure standards

The first standard to be measured is marked on the

display. After the blank value (blank key) measure all
standards (standard key) one by one.
When measuring more than one replicate per
standard, the average value for each standard is
calculated and displayed automatically.
By using the and cursor keys, you also can
select certain standards for measurement. Individual
standards can be remeasured as well.

Softkeys
• [Last cal]: Call up the last saved standard evaluation for this method in order to use it for sample
measurements.
• [Curve fit]: Select the procedure for the standard evaluation. If the result has not been saved, the
method can also be entered later. Instructions for selecting the evaluation procedure can be found in
the Evaluation procedure chapter (see Evaluation with standard curve/line on p. 75).
• [Graph]: Switch to the graphic display of the standard results.
As soon as the minimum number of results for the
evaluation with the selected method (curve fit) is
available, the evaluation result will be shown on the
right side of the display. You can now save the
evaluation and switch to sample measurements via
the [Next >] key.

Graphical view of the standard evaluation.

With the and cursor keys, you navigate between
the standards to view the results. With more than one
replicate per standard, you can switch between the
replicate results using and . You can also select
individual standards from the graphical display and
measure or remeasure them.

Softkeys
• [Table]: Switch to the table display of the standard results.
• [Next >]: Save the standard evaluation and switch to the sample measurement.
 Methods
Eppendorf BioPhotometer® D30 37
English (EN)

6.4.3 Measure samples

The sample key is used for measuring your samples consecutively. Blank results remain saved for one
measuring series, but a new blank result measurement can be performed at any time. With the and
keys you can navigate between the sample results that have been achieved in the measuring series up to
this point.

Results display:
• The concentration result (6 digits with floating
point) is clearly emphasized.
• With graphic: Result to the right of the display.
• Without graphic: Result in the middle of the
display.
• In addition to the result, the basic absorbance
value is shown at a smaller scale.

Additional data
• Upper right; first row:
Sample number: Counted sequentially and reset to "1" for each new series of measurements.
Sample dilution (if provided)
• Upper right; second row:
Sample identification (ID) (if provided)
• Top left:
File name with which the data in the print and export method step can be exported as Excel file (see
p. 43).

Softkeys
• [Dilution]: Enter sample dilution.
• [Edit ID]: Enter sample ID
• [Data]: Display additional result data (not available with all methods).
• [Finish]: End series of measurements and return to method selection.

The displayed absorbance values always correspond to the directly measured values. The
dilution or cuvette factor as well as background absorbances will be incorporated for the
following result calculation (see Absorbance values on p. 73).
 Methods
38 Eppendorf BioPhotometer® D30
English (EN)

Enter dilution

The [Dilution] softkey is activated after the blank

(blank key) has been measured.
1. Press the [Dilution] softkey.
2. Enter the volumes for the sample (up to 3 digits)
and for the dilution buffer (up to 4 digits).
The device will multiply the following sample results
by the calculated dilution factor.

Softkeys
• [Clear dil.]: Delete values for sample dilution.
• [OK]: Confirm sample dilution and return to sample measurement.
• [Cancel]: Cancel entry and return to sample measurement.

The dilution is used for all following sample results until it is changed by a new entry.

Enter sample ID

The ID will be applied to the following sample result. When an ID is being entered the last entered ID will
be displayed as a default template to allow the quick entry of IDs with a consecutive structure. A single ID
can only be assigned once for the same measuring series.

1. Press the [Edit ID] softkey.

2. Enter the sample ID (up to 12 digits).
Alternatives for character input:
• Keypad: If the key is pressed several times in a
row, the possible entries for this key will be
shown consecutively.
• Display keyboard with softkey [Keyboard]: Select
character with the cursor keys and confirm with
enter.

Softkeys
• [Keyboard]: Display keyboard.
• [abc]: Change between upper and lower case letters when making entries with the keypad.
• [OK]: Confirm ID entry and return to sample measurement.
• [Cancel]: Cancel entry and return to sample measurement.
 Methods
Eppendorf BioPhotometer® D30 39
English (EN)

Result image with dilution and ID

Result image with dilution and sample ID

6.4.4 Measure samples: Results displays

This section contains a display of typical results displays for all method groups and an overview of
additional results data, which can be accessed using the [Data] softkey.

Method group Results display Explanation

Absorbance main group
Single λ Results display:
• Absorbance at the measuring
wavelength
• Only for dilution or with cuvettes
other than 10 mm: additional
display of the absorbance value
before the conversion.

Routine main group

 Methods
40 Eppendorf BioPhotometer® D30
English (EN)

Method group Results display Explanation

Nucleic acids Results display:
• Concentration result with
absorbance at the measuring
wavelength
• If activated in the parameters:
Ratios A260/A280
• If activated in the parameters:
Restricted scan.
Navigate between the measuring
points on the graph which are used
for the result calculation with
and .
Additional data ([Data] softkey):.
If the corresponding parameters have
been activated:
• Absorbance value for 280 nm.
• Ratios A260/A230 and absorbance
value for 230 nm.
• Absorbance value for the
background wavelength.
Proteins direct Results display:
UV • Concentration result with
absorbance at the measuring
wavelength
• If activated in the parameters:
Restricted scan.
Navigate between the measuring
points on the graph which are used
for the result calculation with
and .
Additional data ([Data] softkey):.
If the corresponding parameters have
been activated:
• Absorbance value for 260 nm.
• Absorbance value for the
background wavelength.
 Methods
Eppendorf BioPhotometer® D30 41
English (EN)

Method group Results display Explanation

Proteins (with Results display:
reagent) • Concentration result with
absorbance at the measuring
wavelength.
• For evaluation with standard
series: graph of the standard
evaluation with plotted sample
result.

Bacterial density Results display:

• Calculated result with absorbance
at the measuring wavelength.

Basic main group

Factor, standard Analog to Protein direct UV (see above) Results display:
• Concentration result with
absorbance at the measuring
wavelength.
Calibration curve Analog to Proteins (with reagent) (see above) Results display:
• Concentration result with
absorbance at the measuring
wavelength.
• Graph of the standard evaluation
with plotted sample result.
 Methods
42 Eppendorf BioPhotometer® D30
English (EN)

6.4.5 Process results

The sample measurement is followed by two optional steps in the method sequence: process results and
print & export.

In the process results step, you can convert the concentration results to molar concentrations or, after
entering the volumes, into total amounts for the methods of the Nucleic acids group.

As for the result display, you can navigate between the sample results of the measurement series with the
and cursor keys and select results for postprocessing.

Tab. 6-3: More calculations

Press the [More calc.] softkey.

• After the molar mass has been entered (in base/

base pairs or in kDa): Convert the concentration
result to the molar concentration.
• After the sample volume has been entered:
Calculate the total amount in the sample.
• [Save]: Save changes and return to the process
results method step.
• [Cancel]: Cancel and return to the process results
method step.

• For dsDNA the calculation of the molar concentration is based on the assumption of a
double-stranded nucleic acid. For the ssDNA, RNA and Oligo methods, a single-stranded
nucleic acid is assumed.
• For methods which have been reprogrammed via <New Method> in the Routine main
group, Nucleic acids method group, always double-stranded nucleic acids are assumed
for calculating the molar concentration.
 Methods
Eppendorf BioPhotometer® D30 43
English (EN)

After the changes have been saved you can apply

them to all samples of the measuring series with
[Yes].

6.4.6 Print & export

In the last optional method step, you can assemble data packets for all samples of a series of
measurements, or selected samples of a series of measurements, for printing to the printer, export to a USB
stick or export to a PC using a USB cable.

Select data packets

• Use the cursor keys for navigating and confirm
with enter.

Softkeys
• [Print]: Start printing.
• [Export]: Start export.
• [Sample]: Select individual sample results.

Select samples
• Press the [Samples] softkey to call up the sample
selection.
• Use the cursor keys for navigating and confirm
with enter.

Softkeys
• [Select all]: Select all samples
• [De-Sel. all]: Cancel selection.

Data export

The data will be transferred as Excel files (.xls) and can be read with Excel versions Excel 97 and later. For
each of the selected data packets, a worksheet is created in Excel. The file name consists of the method
name, the time and the date of the measuring series.
 Methods
44 Eppendorf BioPhotometer® D30
English (EN)

Select export version

If no USB stick is connected, the first variant cannot

be selected.

Export to USB stick

1. Connect a FAT 32-formatted USB stick to the USB port 4 (see Main illustration on p. 9).
2. Start with [Export] "export to an external storage medium".
Export to PC
Requirement for the PC operating system: Windows XP, SP2 or a higher version.
1. Connect the device to the PC by using the USB cable on the USB port 8 (see Main illustration on p. 9).
2. Prior to beginning a new export make sure that any data that has been exported previously has been
saved to the PC hard drive. Otherwise, the new export will overwrite the data.
3. Start with [Export] "export to PC".
4. The exported data packet will be displayed on your PC as a removable medium named "eppendorf".
Open the Excel file on this drive and save it to the hard drive.

Select data packets

Results Primary result data; cannot be selected because they are always transferred.
Data Additional results data that are displayed during the measurement using the
[Data] softkey.
Graph Restricted absorbance-wavelength-spectrum.
Parameters Method parameters
Standards/results Results data of the standard evaluation.
Standards/graph (Only for standard evaluations with several standards:) absorbance
concentration graph.

Based on the method and parameter setting, only the available data packets are presented.
 Methods
Eppendorf BioPhotometer® D30 45
English (EN)

6.4.7 Finish the series of measurements

After the print & export method step has been finished, you can start a new series of measurements using
the selected method or select a new method.

Finish the series of measurements and start a new series of measurements

• [Next >] softkey: Call up new series method step

• [New] softkey: Call up measure samples method
step and start a new series of measurements.

Finish the series of measurements and select a new method

• [Finish] softkey: Close the series of measurements and call up the method selection.
 Methods
46 Eppendorf BioPhotometer® D30
English (EN)
 Functions
Eppendorf BioPhotometer® D30 47
English (EN)

7 Functions
7.1 Functions of the User main group

With the function key or the [Function] softkey, you reach a menu containing functions like device settings
or calling up saved results.

The functions are structured in 3 columns analog to the method selection. The functions in the User main
group are accessible to you. As for the method selection, you navigate with the cursor keys for selecting
first the desired subgroup and then the desired function in the right column. Press enter to open the
function.

[Info] softkey: call up the firmware version and serial number of the BioPhotometer D30.

Tab. 7-1: Overview of the functions

Subgroup Explanation
Results memory Displays saved results.
The results can be accessed structured according to methods and
measuring series and can be printed and exported directly from the
memory.
General method parameters Parameters which are used for different methods in common are stored
centrally in the Functions area.
Here it is possible to edit (change or create anew) these parameters. In
the Check parameters method step, the comprehensive parameters can
be easily selected using drop-down menus.
• Proteins: Parameters for methods of the Proteins direct UV group
• Units: Units for concentration results which can be used for many
methods.
Absorbance spectra library Absorbance wavelength spectra of important substances, e.g., DNA.
The spectra serve as information and can be used for comparison to a
spectrum of a sample result.
Device settings Editable device settings, e.g., language.
Device calibration • Option for inspecting the photometer. An Eppendorf filter set is
required for this.
Info Open Source Licenses and information on registered trademarks.
 Functions
48 Eppendorf BioPhotometer® D30
English (EN)

7.1.1 Results memory

 In the right column, select the method for which

you would like to call up saved results.
 Confirm with enter.

 Select the desired series of measurement with

the cursor keys.
 Confirm with enter.

As in the method procedure, you can also

successively switch between the display of the
parameters, standards, sample results and, finally,
the data packets for print and export.
The assignment of the softkeys matches the
assignment in the method procedure.
 Functions
Eppendorf BioPhotometer® D30 49
English (EN)

 If you would like to print or export results,

select the data packets.
The procedure for print and export and the
meaning of the function keys corresponds to the
print & export method step.

7.1.2 General method parameters

 In the right column, select the parameter group

for which you would like to edit parameters.
 Confirm with enter.

In this example, the parameter groups for different

proteins are subsumed and stored respectively
under a name. This name can be used to import the
desired parameter group into the method program
when editing a protein method (Proteins direct UV
method group).
Display:
• Left: Name of the protein. Select via and .
• Right: corresponding parameters

Softkeys
• [Edit]: Edit selected parameter group.
• [New]: Create new parameter group.
• [Delete]: Delete selected parameter group.
• [OK]: Return to the function selection.
 Functions
50 Eppendorf BioPhotometer® D30
English (EN)

 To edit a parameter group, use and to

select the parameter which you would like to
edit.
 Confirm with enter.

Softkeys
• [OK]: Save entry and return to the parameter group selection.
• [Cancel]: Return to the parameter group selection without making any changes.

When programming a method of the Proteins direct UV method group, you can access the entries in
General Method Parameter:

Select the name of the protein in order to import

the corresponding parameter group into the
method program. By using the "edit" selection of
the "Nucleic acid" parameter, you also can get
directly to the General Method Parameter function
and view and edit the parameters.

Tab. 7-2: Parameter in General Method Parameter

Parameter Explanation
Proteins When selecting a protein while programming a method of the Proteins
direct UV group, these parameters are loaded into the method
parameters.
• Protein name In order to define a factor for calculating the concentration on the basis of
• Factor the absorbance, enter the following data in addition to the name and
• A0.1% wavelength:
• Ext.coeff. Factor or A0.1% or absorbance coefficient and molar mass.
• Molecular mass
Units You can select a unit from all available units when programming method
parameters.
• Unit Entering a unit that has not yet been programmed for the concentration
result.
 Functions
Eppendorf BioPhotometer® D30 51
English (EN)

• Specifications for proteins which are not preset at the factory can be determined in the
expasy database: http://www.expasy.org/tools/protparam.html.
• A table with A1% values for many proteins can also be found in: C.N.Pace et al., Protein
Science (1995), 4: 2411–2423 (Table 5). The A1% values must be multiplied by 0.1 to
return the required A0.1% values.

7.1.3 Absorbance spectra library

In the right column, you select the spectrum which

you would like to call up and confirm with enter.

Softkeys
• [Export] and [Print]: Export to a USB stick or
print to a PC using a USB cable (see Print &
export on p. 43).
• [OK]: Return to the function selection.

7.1.4 Device settings

The following settings can be modified:

• Language: German, English, French, Spanish,
Italian.
• Date and time.
• Frequency of the automatic self test of the
device after switching on.

Softkeys
• [Save]: Save changes and return to the function
selection.
• [Cancel]: Return to the parameter group
selection without making any changes.
 Functions
52 Eppendorf BioPhotometer® D30
English (EN)

7.1.5 Device calibration

Information on checking the device is provided separately (see Checking the device on p. 54).

7.1.6 Info

The Copyright menu item contains license

information on the Open Source software.
 Maintenance
Eppendorf BioPhotometer® D30 53
English (EN)

8 Maintenance
8.1 Cleaning
DANGER! Electric shock as a result of penetration of liquid.

 Switch off the device and disconnect the power plug before starting cleaning or
disinfection work.
 Do not allow any liquids to penetrate the inside of the housing.
 Do not spray clean/spray disinfect the housing.
 Only plug the device back in if it is completely dry, both inside and outside.

NOTICE! Corrosion from aggressive cleaning agents and disinfectants.

 Do not use corrosive cleaning agents, aggressive solvents or abrasive polishes.

 Do not incubate the accessories in aggressive cleaning agents or disinfectants for a longer
period of time.

1. Wipe down the surfaces with a cloth moistened with a mild cleaning agent.
Cleaning the cuvette shaft
2. The cuvette shaft may only be cleaned with a lint-free cotton swab that has been dampened with ethanol
or isopropanol. Prevent liquid from entering the cuvette shaft. If the shaft needed to be dampened with
water to remove contamination, follow this up by cleaning the shaft with a cotton swab dampened with
ethanol or isopropanol to accelerate the drying process.

8.1.1 Cleaning the cuvette shaft cover

If you would like not only to clean the directly accessible surface of the cuvette shaft cover, you can remove
the cover.

 Do not soak the cuvette shaft cover in cleaning agent.

 Clean the cuvette shaft cover as described.

1. Lift the cuvette shaft cover with one hand.

2. With the other hand, hold the cover at the height
of the locking pin and pull the cover to the right
until the locking pin has been removed.
 Maintenance
54 Eppendorf BioPhotometer® D30
English (EN)

• Pull the cover to the right at a 90 degree angle.

3. Clean the cover with a cloth or lint-free cotton swab dampened with a mild cleaning agent.
4. Slide the locking pin back into the housing as far as it will go.
The locking pin has completely disappeared in the housing.

When the photometer is not being used, close the cuvette shaft using the blue cuvette shaft
cover to protect it from dust and other contamination.

8.2 Disinfection/Decontamination
DANGER! Electric shock as a result of penetration of liquid.

 Switch off the device and disconnect the power plug before starting cleaning or
disinfection work.
 Do not allow any liquids to penetrate the inside of the housing.
 Do not spray clean/spray disinfect the housing.
 Only plug the device back in if it is completely dry, both inside and outside.

1. Clean the device with a mild cleaning agent before the disinfection (see Cleaning on p. 53).
2. Choose a disinfection method which corresponds to the legal regulations and guidelines in place for
your range of application.
3. For example use alcohol (ethanol, isopropanol) or other alcoholic disinfectants.
4. Wipe the surfaces with a cloth which you have moisturized with a disinfectant.
5. If the cuvette shaft cover needs to be removed for the disinfection, proceed as follows for the diassembly
and assembly (see Cleaning the cuvette shaft cover on p. 53).
6. You can use spray disinfection to disinfect the disassembled cuvette shaft cover.

8.3 Checking the device

Requirements
• Observe the ambient conditions (see Ambient conditions on p. 69).
• Perform the check at approx. 20 °C. Prevent temperature changes (e.g. due to open windows).
• Remove the filter from the filter box only for a short time and protect it against contamination or
damages at the filter surfaces.
• Protect the filter against dust, heat, liquids and aggressive vapors.
• The filter is inserted in such a way that the label, with the name of the filter, points towards the detector.
When checking the photometer unit: label shows to the front.
• The cuvette shaft is free from contamination.
 Maintenance
Eppendorf BioPhotometer® D30 55
English (EN)

8.3.1 Checking the photometer unit

Eppendorf offers a filter kit for checking the photometric accuracy and the wavelength systematic error.
The kit contains one blank filter A0 and 3 filters A1, A2 and A3 for checking the photometric accuracy as
well as 2 filters for checking the wavelength systematic error (260 nm, 280 nm). The filter absorbances are
measured in comparison with the blank filter A0. In addition to the information on the accuracy, you also
receive information on the precision: From the respectively 15 measurements per wavelength the variation
coefficient (cv-value) is determined next to the average value.

For measuring, first insert the blank filter (for the blank measurement) and then the test filter like cuvettes
into the cuvette shaft. The absorbance values measured for the test filters are compared with the allowable
value range. For the individual filters, the limit values for the permitted area are printed in a table in the lid
of the filter box.

Please connect the thermal printer if you would like to document the values.

Abb. 8-1: Inner lid of the filter box (model)

BioPhotometer D30 reference filter set

Function : Device calibration/Photometer unit Order No./Best.Nr.: 6133 928.004
Set No./Satz Nr.: 006
Limits measured against Blank A 0 at approx. 20°C
Grenzwerte gemessen gegen Blank A 0 bei ca. 20°C
SN: 6133 914.006 916.006 917.006 921.006 922.006 923.006
Filter Blank Sample Sample Sample Sample Sample
Type A0 260 nm 280 nm A1 A2 A3
Limiting values (A)/Grenzwerte (E)
260 nm 0.000 1.291-1.499 -- 0.197-0.221 1.078-1.101 1.857-1.888
280 nm 0.000 -- 1.041-1.287 0.193-0.217 1.024-1.047 1.717-1.749
320 nm 0.000 -- -- 0.173-0.196 0.923-0.947 1.478-1.509
405 nm 0.000 -- -- 0.161-0.184 0.903-0.926 1.429-1.461
562 nm 0.000 -- -- 0.172-0.195 0.941-0.968 1.496-1.530
595 nm 0.000 -- -- 0.172-0.195 0.943-0.969 1.503-1.538
Random error of wavelength Random error of photometer
Zufällige Messabweichung der Wellenlänge Zufällige Messabweichung des Photometers
Limiting values CV (%)/Grenzwerte VK (%)
260 - 405 nm ≤ 3.0 % ≤ 3.0 % ≤ 2.0 % ≤ 1.5 %
550 - 600 nm ≤ 3.0 % ≤ 3.0 % ≤ 2.0 % ≤ 3.0 %
Wavelength and photometric accuracy/Wellenlängen- und photometrische Richtigkeit
traceable to/rückführbar auf
NIST SRM 2034, SN 04-A und SN 667

Wavelength and photometric characterization of filters:

All characterization is performed on a Cary 100 Bio reference UV-Vis spectrophotometer, SN EL 99023107.
The instrument is requalified regularly by the manufacturer, and is confirmed and documented to
perform within manufacturer's specifications. The current instrument qualification is valid through April 2015.
For the current instrument qualification, the following, NIST traceable, certified secondary spectrometric
calibration standards were used:

Wellenlängen- und photometrische Bestimmung der Filter:

Alle Messungen werden auf einem Cary 100 Bio Referenz-UV-Vis-Spektrophotometer,
Seriennummer EL99023107 durchgeführt.
Dieses Instrument wird regelmäßig vom Hersteller requalifiziert und die spezifikationsgemäße Funktion dokumentiert.
Die aktuelle Qualifizierung ist bis April 2015 gültig und verwendet folgende, NIST-rückführbare und zertifizierte,
spektrometrische Sekundärstandards:

SRM 2031a: SN 667, valid through/gültig bis 12/2014

6133 928.020-00

The valid certificates are part of the requalification documentation.

Die Gültigkeitszertifikate sind Teil der Qualifizierungsdokumentation.

Please protect against dust, heat, cold and liquid.

The limits are valid for max. 2 years. __________________________
Bitte vor Staub, Hitze, Kälte und Flüssigkeiten schützen. Date Signature
Die Grenzwerte gelten für max. 2 Jahre. Datum Unterschrift

Fig. 8-1: Inner lid of the filter box (model)

 Maintenance
56 Eppendorf BioPhotometer® D30
English (EN)

8.3.1.1 Checking the photometric accuracy

1. In the Device calibration group, select the

Photometer unit function and confirm with
enter.

2. Select if you want to check the wavelength

systematic error or the photometric accuracy.
Confirm with enter. Press [Next >] to switch to
the measurement.

3. Follow the instructions on the display and first

measure the blank filter A0 and then the first test
filter A1.
The device will measure the test filter 15 times at
6 wavelengths and then show the average values
as well as the variation coefficients of the series
of measurement for all wavelengths.
 Maintenance
Eppendorf BioPhotometer® D30 57
English (EN)

4. Result display after measuring a test filter for

checking the photometric accuracy.
Measure the other two test filters A2 and A3.

5. Results display after measuring all 3 test filters

for testing the photometric accuracy.
With the and keys, you can view again the
results of the different test filters. Press [Finish]
to complete the test.
6. Compare the average values and variation
coefficients with the supplied table.
If the measured values do not agree with the
permitted range of values, contact Eppendorf
Service.

Have the filter recertified by Eppendorf Service after 2 years.

8.3.1.2 Checking the wavelength systematic error

 To check the wavelength systematic error, proceed as follows: Measure the 3 test filters at the
corresponding wavelength.
 Maintenance
58 Eppendorf BioPhotometer® D30
English (EN)

8.3.2 Device self test

You can set the frequency of the automatic self test (taking approx. 1 minute) with the Device settings
function (see Device settings on p. 51). Ex-factory, the self test interval is set to "weekly".

The self test checks the following items:

• Verification of the detector

– Determination of the random error of the available wavelengths
• Verification of the light source
– The device checks the maximally available light source energy and the light cable quality
– Determination of the random error of a signal on the reference sensor
– Determination of the signal level at the reference sensor
– Separate determination of the light intensity in the UV range
• Determination of the systematic and random error of the wavelength

 In the Device calibration group, select the Perform selftest function and confirm with enter.
After the self test has passed, the display shows the message PASSED.
If the display shows the message FAILED, the self test has failed. If this error cannot be corrected (see
Error messages on p. 62), contact Eppendorf Service.

8.4 Replacing fuses

DANGER! Electric shock.

 Switch off the device and disconnect the power plug before starting maintenance or
cleaning work.

The fuse holder is located between the mains connection socket and the mains power switch.

1 2 3

1. Disconnect the power plug.

2. Press the upper and lower end of the plastic springs 1 together and pull the fuse holder 2 fully out.
3. Replace faulty fuses and reinsert the fuse holder. Make sure that the guiding rail 3 is positioned
correctly.
 Maintenance
Eppendorf BioPhotometer® D30 59
English (EN)

8.5 Decontamination before shipment

If you are shipping the device to the authorized Technical Service for repairs or to your authorized dealer
for disposal please note the following:

WARNING! Risk to health from contaminated device

1. Follow the instructions in the decontamination certificate. You find it as a PDF file on our
website (www.eppendorf.com/decontamination).
2. Decontaminate all the parts you would like to dispatch.
3. Include the fully completed decontamination certificate in the package.
 Maintenance
60 Eppendorf BioPhotometer® D30
English (EN)
 Troubleshooting
Eppendorf BioPhotometer® D30 61
English (EN)

9 Troubleshooting
9.1 General errors

Error Possible cause Remedy

Measuring results are • Reagent is past its shelf  Ensure that the reagent is still within its
imprecise. life. shelf life and properly prepared.
• Reagent has not been  Use clean demineralized water of adequate
prepared properly. quality for preparation if required.
• Pipetting is not correct.  Ensure that the pipette is calibrated and
that pipetting is being performed correctly.
• Incubation procedure  If the method procedure requires
before measurement is incubation before the measurement,
incorrect. ensure that the temperature and time for
incubation are correctly observed.
• The cuvette is  Clean and rinse the cuvette. When
contaminated. replacing a cuvette, pay attention that the
optical window of the cuvette remains
clean and that you do not touch it with your
fingers.
 If the cuvette window has become soiled
from fingerprints, wipe it clean using a
lint-free lab cloth soaked in ethanol or
isopropanol.
• The cuvette is not filled  Ensure that the required minimum volume
completely with of the cuvette for a measurement is
measuring solution, and it reached and that no bubbles are in the
contains bubbles. measuring solution.
• Turbidity of the  Centrifuge the turbid measuring solutions
measuring solution. containing particles and use the clear
supernatant.
• Photometer is drifting.  Contact Eppendorf Service.
 Observe the ambient conditions.
 Prevent temperature changes.
• Cuvette shaft is dirty.  Clean the cuvette shaft.
 Troubleshooting
62 Eppendorf BioPhotometer® D30
English (EN)

Error Possible cause Remedy

The measuring results are • The method has not been  Ensure that the method parameters are
not correct. programmed correctly. entered correctly.
• The standard solution has  Ensure that the correct standard is used
not been prepared and that the measuring solution for the
correctly. standard is prepared correctly.
• The absorbance of the  For instable reagent absorbance and end
reagent is drifting. point methods: When measuring a long
series of samples, measure the reagent
blank value not only at the beginning but
also during the sample series. If the
reagent blank value drifts strongly, the
reagent is not appropriate for error-free
measurements and has to be replaced by a
new reagent.
• The cuvette is not  Position the cuvette in the cuvette shaft so
positioned correctly. that the optical window points towards the
direction of the light path.
 Photometry light path: from back to front

9.2 Error messages

You can exit device displays with error messages using the [OK] softkey.

System errors require an evaluation by the Technical Service. These errors are shown in English (System
error …). Please contact Technical Service in these cases. Other error messages, for which you can carry
out troubleshooting measures, are illustrated in the table below.

Problem Cause Solution

Self test failed. • Cuvette shaft cover was open during  Repeat the self test with empty
self test. cuvette shaft and the cuvette shaft
• The cuvette shaft was not empty cover closed.
during the self test.
• Device is faulty.  Contact Eppendorf Service.
File export failed. During data export:  Reformat or replace the USB stick.
• USB stick improperly formatted or  Reconnect the USB stick and repeat
faulty. the export.
• USB stick removed from the device
too early (during the export).
Failed to initialize • Printer not connected or switched off.  Connect the printer and switch it on.
printer. • Printer not configured correctly.  Reconfigure the printer.
For a correct configuration of the printer
settings refer to the installation
description (see Connecting the printer on
p. 16).
 Troubleshooting
Eppendorf BioPhotometer® D30 63
English (EN)

Problem Cause Solution

Blank • The absorbance of the blank solution  Check the blank solution and
measurement: An used for the blank measurement is too remeasure the blank if required.
intensity on a pixel high.
that influences the • Incorrect or turbid blank solution.
main, auxiliary or
scan wavelength is
too low.
The entered name • Error when entering the name.  See information in the help box.
is not valid. Different causes are possible. For the
precise cause please see the
information in the help box.
A method (or • The name under which the method  Assign a different name.
folder, protein, unit) was saved has already been used for a
with this name different method in the same folder.
already exists. • The message also appears after
edition names already given to a
folder or to a protein or a
concentration unit (under General
Method Parameter).
The following • When opening a method with  Select a different parameter from the
parameter values parameters which access General existing list. If necessary, program a
are not defined in Method Parameter, the system new list entry in General Method
General Method determined that at least one Parameter in order to be able to use it
Parameter: parameter (protein, unit) does not when programming a method.
exist there anymore, so probably has
been deleted.
The value of the This error message appears when editing  Select a different parameter from the
parameter marked method parameters. existing list. If necessary, program a
with * is not defined • Parameter in General Method new list entry in General Method
in the Gen. Param. Parameter is not defined. Parameter in order to be able to use it
Please correct the when programming a method.
parameter.
The entered • See the error text.  Enter the standard concentrations so
standard that the first standard receives the
concentrations are lowest concentration and the other
not monotonically standard concentrations form an
increasing resp. increasing sequence.
monotonically
falling. Correct the
standard
concentrations.
 Troubleshooting
64 Eppendorf BioPhotometer® D30
English (EN)

Problem Cause Solution

At least two of the • See the error text.  Enter the standard concentrations so
entered standard that the first standard receives the
concentrations are lowest concentration and the other
identical. Correct standard concentrations form an
the standard increasing sequence.
concentrations.
The measured • Error when measuring a standard  Repeat the standard measurements or
values are not series: The measured absorbance delete the single, incorrectly
strictly values of the standard series are not measured standard result.
monotonous! continuously increasing or
decreasing.
The ID cannot be • Error when entering the sample ID.  See information in the help box.
set. Different causes are possible. For the
precise cause please see the
information in the help box.
The dilution cannot • Error when entering the dilution.  See information in the help box.
be set. Different causes are possible. For the
precise cause please see the
information in the help box.
There is only one • The number of measurements in one  Start a new series of measurement
measurement left to measuring series is limited to 99. after maximally 99 measurements.
be performed in
this series of
measurement.
The maximum
number of
measurements
within one series of
measurements has
been reached.
 Troubleshooting
Eppendorf BioPhotometer® D30 65
English (EN)

9.3 Result flags

Warnings and error messages for results are displayed in the bottom right of the help box. The header bar
of the Help box is highlighted yellow for warnings and red for error messages.

Warnings: Decide whether the result is useful for you while taking the displayed warning into
consideration.

Error messages: No result is displayed; the reason is shown in the error message.

Problem Cause Solution

The standard curve • No usable result was returned during  Select a different Curve Fit
is not monotone. the evaluation of a standard curve procedure.
Please select using the "spline interpolation",
another Curve Fit. "quadratic regression" or "cubic
regression" Curve Fit procedures.
Some absorbance • For at least one secondary  If the absorbance values of the
values for wavelength, the absorbance exceeded secondary wavelengths are relevant:
secondary the measuring range. Dilute the sample or remove the
wavelengths are too • Secondary wavelengths are not turbidity via centrifugation and repeat
high or are not needed for calculating the the measurement.
displayed. concentration result. They are used
for different purposes. For example,
dsDNA method: absorbance at
280 nm for the calculation of ratios
260/280.
• Turbidity of the measuring solution
• Measurements at the limits of the
photometric measuring range.
The result is • For methods with evaluation via  Accept the measurement result, or
outside the range of standard curves (nonlinear evaluation remeasure the sample under
the standard method): The sample result is up to conditions under which the result is
concentrations. 5 % outside of the standard within the range of the standard
concentration range. concentrations (dilute sample or
modify standard concentrations and
remeasure).
The coefficient of • For methods with evaluation of  Accept the result of the standard
determination is standard series via the regression evaluation or remeasure the
<0.8. procedure: The coefficient of standards.
determination for the regression  Make sure the measuring solutions
evaluation indicates a significant are clear.
deviation of the measuring points
from the regression line.
• Turbidity of the measuring solution.
• Measurements at the limits of the
photometric measuring range.
 Troubleshooting
66 Eppendorf BioPhotometer® D30
English (EN)

Problem Cause Solution

The coefficient of • For methods with evaluation of  Use the sample results with the
determination for standard series via the regression reservation mentioned or repeat the
the regression procedure: If the regression measurement of the standard series
evaluation of the evaluation for the standard series was and samples.
standard series is < nonlinear, but the standard evaluation
0.8. was accepted by the user, a warning
appears after samples have been
measured.
Scan: Some of the • For at least one scan wavelength, the  If the non-displayed areas of the scan
measured absorbance exceeded the measuring are relevant: Dilute the sample or
absorbances are range. remove the dilution via centrifugation
too high and are • Turbidity of the measuring solution. and repeat the measurement.
not displayed. • Measurements at the limits of the
photometric measuring range.
Absorbance at the • Turbidity of the measuring solution.  Measure again considering the
measuring • Optical surfaces of the cuvette are possible causes.
wavelength is too soiled.
high. • Cuvette has been inserted into the
cuvette shaft facing the wrong
direction.
• Too high absorbance of measuring
solution.
The calculated • Measuring solution not prepared  Measure again considering the
result is negative. correctly. possible causes.
• The incorrect factor has been entered
(wrong algebraic sign).
The result has more • Very high sample concentration.  Dilute sample and measure again.
than 6 pre-decimal • Concentration unit does not match  Change the concentration unit
places. the expected range of the sample (Parameter Unit) and measure again.
concentrations.
The result is more • For methods with evaluation via  Remeasure the sample under
than 5 % outside of standard curves (nonlinear evaluation conditions under which the result is
the standard method): within the range of the standard
concentration The sample result is more than 5 % concentrations (dilute sample, modify
range. outside of the standard concentration standard concentrations and
range. remeasure).
• Calculation not • The evaluation required dividing by  Check the reagents and samples used
possible an absorbance result with the value of and repeat the measurement.
because of "zero". This is not mathematically
division by zero. permissible.
Absorbance Examples: Calculation of a factor at
result is zero. one-point calibration; calculation of a
• Calculation 260/280 ratio with nucleic acid
error. Division measurements.
by zero.
 Transport, storage and disposal
Eppendorf BioPhotometer® D30 67
English (EN)

10 Transport, storage and disposal

10.1 Transport

 Use the original packaging for transport.

Air temperature Relative humidity Atmospheric pressure

General transport -25 °C – 60 °C 10 % – 95 % 30 kPa – 106 kPa
Air freight -40 °C – 55 °C 10 % – 95 % 30 kPa – 106 kPa

10.2 Storage

Air temperature Relative humidity Atmospheric pressure

In transport packaging -25 °C – 55 °C 25 % – 75 % 70 kPa – 106 kPa
Without transport -5 °C – 45 °C 25 % – 75 % 70 kPa – 106 kPa
packaging

10.3 Disposal

In case the product is to be disposed of, the relevant legal regulations are to be observed.

Information on the disposal of electrical and electronic devices in the European Community:

Within the European Community, the disposal of electrical devices is regulated by national regulations
based on EU Directive 2012/19/EU pertaining to waste electrical and electronic equipment (WEEE).

According to these regulations, any devices supplied after August 13, 2005, in the business-to-business
sphere, to which this product is assigned, may no longer be disposed of in municipal or domestic waste. To
document this, they have been marked with the following identification:

Because disposal regulations may differ from one country to another within the EU, please contact your
supplier if necessary.
 Transport, storage and disposal
68 Eppendorf BioPhotometer® D30
English (EN)
 Technical data
Eppendorf BioPhotometer® D30 69
English (EN)

11 Technical data
11.1 Power supply

Power supply 100 V to 240 V ±10 %, 50 Hz to 60 Hz

Overvoltage category II
Degree of pollution 2
Power consumption Maximum power consumption according to name plate: 25 W
Approx. 15 W during operation
Approx. 5 W with the display dimmed
Permitted mains interruption Approx. 10 ms at 90 V
Approx. 20 ms at 230 V
Protection class I
Fuses T 2.5 A/250 V, 5 mm × 20 mm (2 pcs.)

11.2 Ambient conditions

Operation Ambient temperature: 15°C to 35°C

Rel. humidity: 25% to 70%
Air pressure: 86 kPa to 106 kPa
Air pressure Use up to an altitude of 2000 m above MSL

Do not expose to direct sunlight.

11.3 Weight/dimensions

Weight 5.4 kg
Dimensions Width: 295 mm
Depth: 400 mm
Height:150 mm
Space required Width: 500 mm (with thermal printer: 750 mm)
Depth: 500 mm
 Technical data
70 Eppendorf BioPhotometer® D30
English (EN)

11.4 Photometric properties

Measuring principle Single beam absorption photometer with reference beam
Light source Xenon flash lamp
Monochromator Holographic aberration-corrected concave grating
Beam receiver CMOS photodiodes
Wavelengths 230 nm, 260 nm, 280 nm, 320 nm, 340 nm, 405 nm, 490 nm, 562 nm,
595 nm, 600 nm
Wavelength selection Method-dependent, freely selectable
Spectral bandwidth ≤ 4 nm
Systematic wavelength error ±1 nm
Random wavelength error ≤ 0.5 nm
Photometric measuring range 0 A to 3.0 A at 260 nm
Reading accuracy ΔA = 0.001
Random photometric error ≤ 0.002 at A = 0
≤ 0.005 (0.5 %) at A = 1
Systematic photometric error ±1 % at A = 1
Stray light component < 0.05 %

11.5 Further technical parameters

Cuvette material For measurements in the UV:
Quartz glass or UV transparent plastic (Eppendorf UVette, 220 nm to
1600 nm)
For measurements in the visible range:
Glass or plastic material
Overall cuvette height Min. 36 mm
Height of the light beam in 8.5 mm
the cuvette
Key pad 22 foil keys
6 foil keys as softkeys
Result output Absorbance, concentration, restricted scan (absorbance wavelength
spectrum)
Additional, method-dependent data (ratio, background absorbances)
Display VGA TFT display 5.7”
Operator guidance language English, French, Spanish, Italian, German
Interfaces USB master: for USB stick and thermal printer DPU-S445
USB slave for connecting to a PC
Serial interface RS 232: for thermal printer DPU-414
Connected devices must meet the safety requirements specified in IEC
60950-1.
 Technical data
Eppendorf BioPhotometer® D30 71
English (EN)

11.6 Application parameters

Methods Preprogrammed and freely programmable methods for all measuring and
evaluation procedures:
• Absorbance measurements
• Nucleic acids and proteins, OD600
• Methods with evaluation via factor, standard and standard series
Method-dependent Absorbance, concentration via factor and standard.
evaluation Concentration via standard series:
• Linear regression
• Nonlinear regression (2nd and 3rd degree polynoms)
• Spline evaluation
• Linear interpolation (point-to-point evaluation)
Additional data for nucleic acids: Ratios 260/280 and 260/230; molar
concentration, total yield
Method memory >100 method programs
Measured value memory and Memory for >1 000 results with all data of the results evaluation and
calibration memory standard evaluation, sample number, sample name, date and used
parameter set of the method program.
(The number of saved results depends on the number of saved methods.)
 Technical data
72 Eppendorf BioPhotometer® D30
English (EN)
 Evaluation procedure
Eppendorf BioPhotometer® D30 73
English (EN)

12 Evaluation procedure

This chapter describes the evaluation procedures available in the method programs as well as the
calculation of a dilution using the device software.

When comparing the measuring results to the results of other photometers/

spectrophotometers, note that the values may be dependent on the bandwidth of the devices.
In the following cases the differences may be significant:
• The absorbance spectrum shows a narrow peak in the measurement wavelength.
• The measurement is carried out not at the maximum but at the edge of a peak.
Therefore, check the accuracy of the methods by measuring standards.

12.1 Absorbance values

Absorbance values are displayed as AXXX (XXX represents the wavelength). These displays always match
the directly measured values, i.e., without corrections, which are incorporated in the final evaluation, e.g.,
corrections for optical path lengths of the cuvette, or background corrections.

12.1.1 Blank

All absorbance values are always related to the last measured blank (blank). Therefore, a blank
measurement is compulsory at the start of every series of measurements and can be completed at any time
during a series of measurements. Ideally, the blank measurement should be able to compensate for any
influences on the absorbance value of the measuring solution. The blank should therefore be measured
with the same buffer that was used for the sample measurement and the same cuvette that was used to
measure the sample value – unless the cuvettes used for the blank and sample measurements are optically
aligned and thus have the same absorbance value at the measuring wavelength.

12.1.2 Background correction

Main application: Partial correction of distortions of the absorbance for nucleic acid measurements due to
turbidity in the measuring solution. For example, the absorbance at 320 nm, which should be approx. 0 A
with pure nucleic acids, is subtracted from the absorbance at 260 nm, (the measuring wavelength for
nucleic acids).

AXXX ,corrBkgr AXXX  ABkgr

AXXX, corrBkgr = calculated corrected absorbance at a wavelength of XXX nm.

AXXX = measured absorbance at a wavelength of XXX nm.

ABkgr = measured absorbance at the background wavelength.

 Evaluation procedure
74 Eppendorf BioPhotometer® D30
English (EN)

12.1.3 Cuvette correction

All absorbance values which are used for result calculation are standardized to the cuvette layer thickness
of 10 mm. If a cuvette with a different path length is used, this path length must be defined in the cuvette
parameter. In this case, the measured absorbances are corrected to match measuring results with a cuvette
layer thickness of 10 mm before converting them to sample results.

This correction is applied to:

• Methods with evaluation by factor.
• Methods of the Absorbance group, for which only absorbance values are output

The correction is not applied to:

• Methods with evaluation by standards, as we presume that standards and samples are measured in
cuvettes of the same layer thickness.
• Calculation of ratios as A260/A280 (for nucleic acid measurements).

10
AXXX ,corrCuv AXXX u
Cuv

AXXX, corrCuv = calculated corrected absorbance at a wavelength of XXX nm.

AXXX = measured absorbance at a wavelength of XXX nm.

Cuv = path length of the cuvette.

12.2 Evaluation with factor or standard

C Au F

C = calculated concentration.

A = absorbance.

F = factor.

The factor is programmed in the parameter list and can be modified. It always relates to an optical path
length of the cuvette of 10 mm. If you change the Cuvette parameter the device will take the modification
into account when calculating the results. Therefore you do not need to change the factor for the
evaluation.

If, on the other hand, you modify the concentration unit, you have to ensure that the factor is adjusted for
the selected unit.
 Evaluation procedure
Eppendorf BioPhotometer® D30 75
English (EN)

The factor is either entered directly as a parameter during the "Factor" evaluation procedure or calculated
during the "Standard" evaluation procedure (evaluation with a standard concentration):

CS
F
AS

F = calculated factor

CS = concentration of the standard (enter as parameter).

AS = measured absorbance of the standard.

If multiple measurement (2 or 3 replicates) has been programmed for the standard, the average value is
calculated from the measured absorbance values and inserted as AS.

12.3 Evaluation with standard curve/line

If evaluations are made with more than one standard, the following evaluation procedures for the standard
curve/line can be selected with the [Curve fit] in the measure standards/new method step:

Evaluation procedure Description Minimum required number of

standard points
Linear interpolation Linear point-to-point connection 2 standards minimum.
in the absorbance concentration
graph of the standard evaluation.
Linear regression Polynome regression for first 3 standards minimum.
degree polynomial.
Quadratical regression Polynome regression for second 4 standards minimum.
degree polynomial.
Cubical regression Polynome regression for third 5 standards minimum.
degree polynomial.
Spline interpolation Interpolation via natural cubic 3 standards minimum.
splines.

For the regression procedure, one can select that the regression line (regression curve) goes through the
zero point.
 Evaluation procedure
76 Eppendorf BioPhotometer® D30
English (EN)

• Use the "linear regression" procedure for calibration lines.

• With curvilinear gradients, test which evaluation procedure (quadratic regression, cubic
regression, spline interpolation) produces the function that is most suitable to the standard
evaluation. Spline interpolation connects the measuring points by cubic polynomials,
whereas the regression methods position a quadratic or cubic function between the
measuring points in such a way, that the smallest possible deviation from the function
results for the measuring points.
• Aside from the calculated regression equation, the regression method also displays the
coefficient of determination as a measure for the scattering of the measuring points around
the calculated function. At a value of < 0.8 for the coefficient of determination the result is
issued with a warning.
• If the first standard hat a concentration of "0", select the setting in which the regression
line (regression curve) goes through the zero point.
• If none of the procedures recommended for curvilinear gradients produce satisfactory
results, select the "linear interpolation" procedure.

12.4 Dilution

In the measure samples method step. entered dilutions are considered in the result calculation:

CDil, corr = result converted using the dilution factor

VS = volume of the sample in the measuring solution

VDil = volume of the diluent in the measuring solution

 Evaluation procedure
Eppendorf BioPhotometer® D30 77
English (EN)

12.5 Special evaluation procedures for nucleic acids and protein UV

This section covers the evaluation of nucleic acids or proteins in the Nucleic acids and Proteins direct UV
method groups.

12.5.1 Ratios A260/A280 and A260/A230.

Application: Information on the purity of the measured nucleic acid. Application of the evaluation
procedure can be activated in the A260/280 or A260/A230 parameters.

"Ratio" refers to the quotients of the measured absorbances at the listed wavelengths.

Literature values for ratio values with pure nucleic acids:

A260/A280
• DNA: 1.8 to 1.9
• RNA: 1.9 to 2.0
(Current Protocols in Molecular Biology, 1994)

A260/A230

For the ratios A260/A230, different information can be found in the literature for pure nucleic acids:
• DNA: 2.3 to 2.5
(The Nucleic Acids, 1955)
• DNA: 1.9
(Current Protocols in Molecular Biology, 1994)

The values are highly dependent on the pH value. Therefore, nucleic acids should not be measured in
water, but in a buffer with a pH of 7 to 7.2 (e.g., TE buffer).

12.5.2 Conversion to molar concentrations and nucleic acid quantities

The conversion only can be applied to nucleic acids. It is realized in the process results/More
calculationsmethod step.

12.5.2.1 Calculation of amount

Application: calculating the amount (mass) of nucleic acid in the total sample volume.

M = calculated total amount (mass) of nucleic acid in the sample tube. Unit: μg.

C = nucleic acid concentration calculated from the measurement. Unit: μg/mL or ng/μL.

VS, total = total volume of the sample in the sample tube. Enter this value in More calculations. Unit: μL.
 Evaluation procedure
78 Eppendorf BioPhotometer® D30
English (EN)

12.5.2.2 Calculation of the molar concentration

Application: calculation of the molar concentration of the nucleic acid from the mass concentration and
relative molar mass. The molar mass is either entered directly or calculated by the device from the entered
number of bases or base pairs per nucleic acid molecule.

C u 103
CMol
MM

CMol = calculated molar concentration of the nucleic acid. Unit: pmol/mL.

C = nucleic acid concentration calculated from the measurement. Unit: μg/mL or ng/μL.

MM = relative molar mass. Unit: kDa

If the number of bases or base pairs per nucleic acid molecule are entered in More calculations instead of
the relative molar mass, the MM is calculated from the number of the bases or base pairs:

For dsDNA:

For ssDNA, RNA, Oligo:

MM = calculated relative molar mass; unit: kDa

bp = entered number of base pairs per molecule

b = entered number of bases per molecule

• For dsDNA the calculation of the molar concentration is based on the assumption of a
double-stranded nucleic acid. For the ssDNA, RNA and Oligo methods, a single-stranded
nucleic acid is assumed.
• For methods which have been reprogrammed via <New Method> in the Routine main
group, Nucleic acids method group, always double-stranded nucleic acids are assumed
for calculating the molar concentration.
 Evaluation procedure
Eppendorf BioPhotometer® D30 79
English (EN)

12.5.3 Calculating the factor for protein in "General Method Parameter"

This section only covers the calculation of the protein component in the Proteins direct UV method group.
For this method group, the protein component is selected in the parameters (see Method parameters on
p. 32). The protein component is assigned a factor that will be entered in the General Method Parameter/
Proteins function for each protein. Alternatively, A0.1% or the absorbance coefficient plus the molar mass
of the protein can be entered instead of the factor. In this case, the factor is calculated as follows:

1
FP
A0.1%

F = factor for the protein; unit: g/L.

A0.1% = absorbance of the protein at a concentration of 0.1 % (1 g/L).

When entering the molar absorbance coefficient and the relative molar mass of the protein A0.1% can be
calculated on this basis:

HP
A0.1%
MM P

εP = molar absorbance coefficient of the protein; unit: cm-1M-1.

MMP = relative molar mass of the protein; unit: Da (entry in General Method Parameter in kDa).
 Evaluation procedure
80 Eppendorf BioPhotometer® D30
English (EN)
 Ordering information
Eppendorf BioPhotometer® D30 81
English (EN)

13 Ordering information
Order no. Order no.  Description
(International) (North America)
Eppendorf BioPhotometer D30
6133 000.001 – 230 V / 50 – 60 Hz, mains/power plug Europe, more types of
mains/power connection available
6133 000.010 6133000010 120 V / 50 – 60 Hz, mains/power plug North America
Eppendorf μCuvette G1.0 and BioPhotometer D30 (bundle)
Eppendorf microvolume measuring cell and BioPhotometer
D30
6133 000.907 6133000907 230 V / 50 – 60 Hz
6133 000.908 6133000908 120 V / 50 – 60 Hz
BioPhotometer D30 reference filter set
6133 928.004 6133928004 Filter set for checking photometric precision and wavelength
accuracy (according to NIST)
Thermal Printer DPU-S445
including power supply and printer cable
6135 011.000 230 V, EU
6135 010.004 6135010004 115 V/110V, USA, JP
6135 012.007 230 V, UK
Thermo paper
0013 021.566 952010409 5 rolls
Eppendorf μCuvette G1.0
6138 000.018 6138000018 Eppendorf microvolume measuring cell for Eppendorf
BioPhotometer and BioSpectrometer
Eppendorf UVette 220 nm – 1 600 nm
Original Eppendorf plastic cuvette, PCR clean, Protein-free
0030 106.300 952010051 50 - 2 000 μL, 80 pieces, individually packaged
Eppendorf UVette routine pack 220 nm – 1 600 nm
Eppendorf Quality
0030 106.318 952010069 50 - 2 000 μL, 200 pieces, reclosable box
Eppendorf macro Vis Cuvettes
0030 079.345 0030079345 10 × 100 pieces
Eppendorf semi-micro Vis Cuvettes
0030 079.353 0030079353 10 × 100 pieces
Cuvette stand
4308 078.006 940001102 for 16 cuvettes
 Ordering information
82 Eppendorf BioPhotometer® D30
English (EN)
Certificates
Evaluate your manual

Give us your feedback.

www.eppendorf.com/manualfeedback

Your local distributor: www.eppendorf.com/contact

Eppendorf AG · 22331 Hamburg · Germany
eppendorf@eppendorf.com · www.eppendorf.com