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Limiting Dilution Protocol

Notes:

Limiting dilution analysis attempts to determine the frequency of cells having a particular function that are present in a mixed population of cells. So limiting dilution analysis is an easy-operated method that used to measure the abundance of cells able to perform a particular function.

Procedure:

Day before limiting dilution:

1. Refresh the culture medium

Day of limiting dilution:

1. Collet all cells

2. Count out the number and estimate the viability (should be higher than 80%)

3. Calculate the dilutions to 4cells/well, 2cells/well and 1cell/well, then serially diluted to designated concentration

4. Pour each of dilutions into 96-well plate and culture in incubator till colonies could be observe

For research use only. Not intended for any clinical use.

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