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.