Cell counting/plating
Overview
Count the number of viable cells in a culture via dilution plating. The following assumes E. coli cells but it should apply to any type of cells.
Materials
LB agar plate
LB media for dilution
8-tube strips (optional but easiest)
Procedure
Fill each tube in the dilution with 90 μl of LB
Add 10 μl of the sample to the first tube and mix
From the first tube, remove 10 μl and mix into second tube
Repeat for the number of dilutions you wish to do (8 should be more than enough)
Take 10 μl from each dilution and spot it on to the agar plate
Allow droplet to dry and incubate
The first dilutions will contain a thick lawn of cells and the last dilutions will contain no cells. There should be one drop which contains countable single colonies. From this, you can calculate the number of cells in the original sample. For example, if there 4 colonies on dilution 5, there are 4E4 cells/μl.
Count the number of viable cells in a culture via dilution plating. The following assumes E. coli cells but it should apply to any type of cells.
Materials
LB agar plate
LB media for dilution
8-tube strips (optional but easiest)
Procedure
Fill each tube in the dilution with 90 μl of LB
Add 10 μl of the sample to the first tube and mix
From the first tube, remove 10 μl and mix into second tube
Repeat for the number of dilutions you wish to do (8 should be more than enough)
Take 10 μl from each dilution and spot it on to the agar plate
Allow droplet to dry and incubate
The first dilutions will contain a thick lawn of cells and the last dilutions will contain no cells. There should be one drop which contains countable single colonies. From this, you can calculate the number of cells in the original sample. For example, if there 4 colonies on dilution 5, there are 4E4 cells/μl.
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