I tried to prepare electro competent cells with 10% glycerol. I found it difficult to pellet them down properly. The cells adhere very loosely to the walls of tubes after centrifugation. This caused loss of cells during washing. I used 5000 rpm for 10 mins.
Any suggestion to avoid cell loss during the whole process?
Fereshteh Sadat Younesi
you're rpm is so low. you increase rpm (14000) or time ( in case of time, you should be sure about the cold temperature)
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Gaurav Kumar
Thanks Fereshteh for the suggestion, but raising rpm to 14000 will make the cells more vulnerable to lysis as they become more fragile.
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Fereshteh Sadat Younesi
for heat shock I use 13000 rpm. if you think it is possible that high rpm make cells more fragile, you can increase your time.
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