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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.
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