Hi,
I need to perform colony transfection assay follwing transfection of my gene of interest (GOI). After transecting my cells for 24hrs, how many cells do I seed in a 6 well plate for colony formation? 500 or 2000? Any ideas? I plated 15000 cells over 2 weeks and it was not possible to conunt them after crystal violet staining. Any recommendations to the procol will be appreciated.
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amelia417
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Posted 09 March 2010 - 09:32 AM
I seed 500 or 1000 cells of moderately quickly growing liver cancer cell lines in 6-cm dishes for about 14 days. Depending on the growth rate of your cells, and since you are using 6-well plates with a smaller surface area, I would recommend no more than 500 per well to start.
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SF_HK
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Posted 09 March 2010 - 09:10 PM
amelia417, on Mar 10 2010, 01:32 AM, said:
I seed 500 or 1000 cells of moderately quickly growing liver cancer cell lines in 6-cm dishes for about 14 days. Depending on the growth rate of your cells, and since you are using 6-well plates with a smaller surface area, I would recommend no more than 500 per well to start.
Thank you for the suggestion. I'm working with liver cancer cell lines too and I'm planning to seed PLC and 97L cells, both of which grow fast and thus 500 cells should be good.
