I have been trying to do Clonogenic assay for a while now. It has been unsucessful so far standardizing the right number of cells to form colony in a 6 well plate.
Kindly guide me in preparing this
Manzoor Ahmed
Count the total number of cells by using a haemocytometer. Use the formula ( required no. of the cell (i.e 500) / Stock (i.e total number of cells) * volume as per your requirement.
E.g, The total no. of cells was 1,00,000 and we need two ml media volume per well. So for 6 wells total volume would be 12 ml.
It can be calculated as:
500/100000* 12ml
0.06 ml or 60 microlitres of cell suspension and 11940 microlitres of media.
Mix well by pipetting and add 2 ml media to each well.
In this way, you will get 500 cells/well.
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