Scribd
Upload
14 views3 pages

206 Lab Ex - 5 - Plate Isolation Techniques

The document outlines Lab Lesson 5 on the plate technique for isolating pure cultures in microbiology. It describes methods such as the streak plate, spread plate, and pour plate techniques, emphasizing the importance of isolating single bacterial colonies for study. The procedure includes steps for preparing agar plates, inoculating them, and incubating to observe growth patterns.

Uploaded by

rolandoosorio1230
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
14 views3 pages

206 Lab Ex - 5 - Plate Isolation Techniques

The document outlines Lab Lesson 5 on the plate technique for isolating pure cultures in microbiology. It describes methods such as the streak plate, spread plate, and pour plate techniques, emphasizing the importance of isolating single bacterial colonies for study. The procedure includes steps for preparing agar plates, inoculating them, and incubating to observe growth patterns.

Uploaded by

rolandoosorio1230
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 3

MICROBIOLOGY AND PARASITOLOGY Laboratory

Lab Lesson 5
Plate Technique of Isolating Pure Culture
In the natural setting microbes exist as one part of a complex community of differing
population of organisms. Isolation techniques are applied to separate certain type of bacteria
that enable one to study it morphological, cultural and physiological characteristic. After
isolating a certain colony, this can be grown as pure culture. Pure cultures are comprised of
one species of bacteria. This culture possible be kept or maintained in a culture collection.
Several methods are employed in obtaining pure culture. Each has certain advantages
and limitations. Any method used in isolating microbes must ensure that a single cell is
introduced into a sterile medium using appropriate culture glasswares.
Plating is the most common technique used in isolating pure culture. On a sterile
medium a sample is placed at one site and then dispersed on the growth medium. As the
sample is scattered, cells become isolated. Each isolated cell will produce a colony on the
culture medium.
Three common plating methods used routinely in the laboratory include pour plate,
streak plate, and spread plate.

a) Streak Plate Method


A small amount of sample is transferred using either a loop or needle on the surface
of a suitable, agar medium either by loop or transfer needle. Successive streaks are
made to provide dilutions that eventually produce isolated colonies.

b) Spread Plate Method


A known diluted sample is placed in the agar surface and is scattered evenly using a
bent glass rod.

c) Pour Plate Method


One ml each of successively diluted samples is mixed with 9 ml of sterile, molten
agar maintained at 450C in a petri dish. The contents are carefully mixed and cooled to
solidify. The dishes are incubated at 350C.

Activity
Objective
At the end of the exercise the students must be able to isolating pure culture using
streak plate method.

Materials

By pair:
4 agar plates
4 inoculating loops
4 alcohol lamps
Mixed bacterial culture in broth tubes

19
Lab Lesson 5: Plate Techniques of Isolating Pure Cultures
MICROBIOLOGY AND PARASITOLOGY Laboratory

Procedure
1. Prepare your table by disinfecting its surface with alcohol available.
2. Label the bottom surface of your agar plate with your name, section and date.
3. With a sterilized loop obtain mixed culture of bacteria by dipping the loop into the culture
tube.
4. Streak each plate with one of the methods described below.

Streak Plate Technique

Quadrant Streak Method


a. Spread the organism over a small area near the edge of the plate. This is area 1
(Figure 5-1).
b. Apply the loop lightly to the medium to avoid
digging into it.
c. Flame the loop and cool it for 5 seconds.
d. Make five or six streaks from area 1 to area 2.
e. Flame the loop again and allow it to cool.
f. Make five or six streaks from area 2 to area3
g. Flame the loop again and make five or six streaks
from area 3 to area 4.
h. Flame the loop again before putting it down.

Figure 5-1. Quadrant streak method of


obtaining pure culture.

Radiant Streak Method


a. Spread the organism over a small area near the
edge of the plate (5-6 streaks). This is area 1
(Figure 5-2).
b. Flame the loop and allow it to cool for 5 seconds.
c. From the edge of area 1 make 7 or 8
streaks to opposite side of the plate.
d. Flame the loop again and cross streak over the last
group of streaks (7 or 8 streaks).
e. Flame the loop again before putting it down.

Figure 5-2 Radiant streak method.

5. Incubate plates at 35O C for 24-48 hours. Inoculated plates are always incubated in an
inverted position.
6. Look for isolated colonies. Draw the colonies.

20
Lab Lesson 5: Plate Techniques of Isolating Pure Cultures
MICROBIOLOGY AND PARASITOLOGY Laboratory

Name: _____________________________ Score: ____________________


Lab Schedule: _______________________ Term/Date: ___________________

Lab Report 5 Plate Techniques of Isolating Pure


Cultures
Attendance Stamp
Results and Observation
Examine each of the streak plates, and make a sketch
indicating the distribution of growth.

Streak Plate Method


Compare the two patterns.

---------------------------------------------------------------------------------------------------------------------
---------------------------------------------------------------------------------------------------------------------

Radiant streak Quadrant streak

Questions

1. Why is the loop being sterilized before transferring from one area to another area?

2. Why is serial dilution of inoculum important in obtaining pure culture?

3. Which area in the radiant streak obtained isolated colonies?

….. quadrant streak?

4. Where do you observe the growth (surface, subsurface or bottom)?


Why is this so?

21
Lab Lesson 5: Plate Techniques of Isolating Pure Cultures

You might also like

pFad - Phonifier reborn

Pfad - The Proxy pFad of © 2024 Garber Painting. All rights reserved.

Note: This service is not intended for secure transactions such as banking, social media, email, or purchasing. Use at your own risk. We assume no liability whatsoever for broken pages.


Alternative Proxies:

Alternative Proxy

pFad Proxy

pFad v3 Proxy

pFad v4 Proxy