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Tricks for making competent cells of difficult strains

Tricks for making competent cells of difficult strains - Molecular Cloning Forum

Tricks for making competent cells of difficult strains - DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.


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Old 09-12-2010, 09:44 PM
Pipette Filler
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Default Tricks for making competent cells of difficult strains



I want to use a knockout E. coli strain for my protein expression. In the literature, it has been used before, but with electroporation. We have had no luck making the cells chemically competent and I do not have access to an electroporator.

Does anyone have any suggestions for getting tricky strains to take up DNA? Here is the genotype, in case it matters:
thr1 araC14 leuB6(Am) lacY1 glnV44(AS) galK2(Oc) λ- trp-59 mdh-2 rpsL265(strR) xylA5 mtl-1 thi-1
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