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Serial Dilution: Course: MCB 103 Lab Section: 1 Name: Noor-E-Khadiza Shama ID: 1921168 Date:10/03/20

The document describes an experiment on serial dilution. Serial dilution aims to reduce the number of bacterial colonies in plates so they can be easily counted to calculate the initial concentration. The experiment involves transferring 1 ml of sample (tamarind juice) into test tubes containing 9 ml of saline, creating 10-fold dilutions up to 10-4. This results in dilution factors of 1:10, 1:100, 1:1000, 1:10000 respectively. The volume of original sample decreases by a factor of 10 with each dilution, and the bacterial concentration is expected to decrease similarly. Precautions for the micropipette and contamination prevention are outlined.

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160 views3 pages

Serial Dilution: Course: MCB 103 Lab Section: 1 Name: Noor-E-Khadiza Shama ID: 1921168 Date:10/03/20

The document describes an experiment on serial dilution. Serial dilution aims to reduce the number of bacterial colonies in plates so they can be easily counted to calculate the initial concentration. The experiment involves transferring 1 ml of sample (tamarind juice) into test tubes containing 9 ml of saline, creating 10-fold dilutions up to 10-4. This results in dilution factors of 1:10, 1:100, 1:1000, 1:10000 respectively. The volume of original sample decreases by a factor of 10 with each dilution, and the bacterial concentration is expected to decrease similarly. Precautions for the micropipette and contamination prevention are outlined.

Uploaded by

Shama
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Course: MCB 103 Lab

Section: 1
Name: Noor-E-Khadiza Shama
ID: 1921168
Date:10/03/20

Experiment 8

SERIAL DILUTION
PURPOSE
Serial dilution is usually done before plating. It aims to reduce the number of bacterial colonies
in plates so that they can be counted easily and the initial concentration of cells in the original
culture can be calculated

PRINCIPLE
In 10 fold serial dilution ,when 1ml original culture is transferred to 9ml saline in a test tube
“A”, the culture will be diluted 10 times or we will get 1:10 dilution. If we then transfer 1 ml
solution from test tube “A” into test tube “B” having 9ml saline, we will get 100 times diluted
solution of the original culture. If we further dilute the solution of tube “B”,we will get 1000
times diluted solution of the original culture.

MATERIALS & EQUIPMENT


1. 4 test tubes
2. A test tube rack
3. Saline solution
4. A Micropipette
5. Sample solution (tamarind juice)
6. Vortex mixer
7. Bunsen burner

PROCEDURE
1. 4 tubes are taken and each tube is labelled with 10-1 ,10-2 ,10-3 and 10-4
2. Each tube is filled with exactly 9 ml of saline solution
3. A micropipette is used to transfer 1 ml of original culture (tamarind juice) in the 10 -1 test
tube containing 9 ml saline. So a total of 10 ml solution will be formed after transferring,
dilution factor will be 1:10
4. Then the micropipette is used to transfer 1 ml solution from 10 -1 test tube in to 10-2 test
tube forming a total of 10 ml solution with dilution factor 1:100
5. Then the micropipette is again used to transfer 1 ml solution from 10 -2 test tube in to 10-
3
test tube, this time the dilution factor will be 1:1000
6. Finally, the micropipette is used to transfer 1 ml solution from 10 -3 test tube in to 10-4
test tube forming a total of 10 ml solution and the dilution factor is 1:10000
7. Discard 1ml of solution from the last test tube
8. Micropipette caps are changed after transferring solution each time.
9. After transferring each time, the micropipette is pushed gently to rinse the solution in
the test tube
10. Before transferring the solution each time, it is mixed evenly using a vortex mixture and
the mouth of the tube is burned in the Bunsen burner flame.

RESULT & OBSERVATION

TUBE DILUTION FACTOR VOLUME OF ORIGINAL SAMPLE


First tube 10 -1
1:10 1 ml
Second tube 10-2 1:100 0.1 ml
Third tube 10-3 1:1000 0.01 ml
Forth tube 10-4 1:10000 0.001 ml

DISCUSSION
In the result, we can see that after 10 fold dilution each time, the volume of original sample is
decreasing by a factor of 10. So we can say that with each dilution, the concentration of
bacteria also decrease by factor 10. If there are 100 bacteria in the first tube ,we expect to get
10 bacteria in the second tube and so on.

PRECAUTION
1. The micropipette should not be turned upside down after taking in the solution
2. Tips should be changed each time to prevent contamination of one solution by another
3. Pressing the micropipette too hard should be avoided
4. The tip of the micropipette should not touch anywhere during the process
5. Careful measures must be taken while handling electronic devices like vortex mixture

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