Dear all Molecular Biologist,
I am trying to prepare competent cell for transformation. but i am confused to use glycerol or DMSO as a cryopretectant. there is any chance to reduce/increase transformation efficiany by using Glycerol or DMSO, And which one is best for storage of Ecoli competent cell.
Could anyone shed light on this problem.
I think the competence of your cells depends on the prep method rather than the final cryoprotectant. For example in Chung's DMSO method, DMSO is used as a cryoprotectant since it is already in the buffer. Same is true for CaCl2 method in which glycerol is part of the buffer. I do not know what method you are using or what competence you require but the majority of the most common prep methods contain glycerol in the competence buffer thus it should be fine. I think both should be fine unless you are going to electrocompetent cells then it might be better to use glycerol as the cryoprotectant instead of DMSO.
I think the competence of your cells depends on the prep method rather than the final cryoprotectant. For example in Chung's DMSO method, DMSO is used as a cryoprotectant since it is already in the buffer. Same is true for CaCl2 method in which glycerol is part of the buffer. I do not know what method you are using or what competence you require but the majority of the most common prep methods contain glycerol in the competence buffer thus it should be fine. I think both should be fine unless you are going to electrocompetent cells then it might be better to use glycerol as the cryoprotectant instead of DMSO.
Hi,
I used 15-30% glycerol as cryoprotectant for my E. coli competant cells. It worked for me.
Dear Kamal,
The steps that you use for making competent cells will lead you to your final storing solution and it works fine. Be it glycerol, PEG or whichever method you are following. Eventually with time the competence will decrease, but glycerol works fine. Ultimately when you use your competent cells, the storing solution should not hinder your transformation process. Best Wishes.
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