competent cell preparation

Sasha Kraev kraev at bc.biol.ethz.ch
Wed Nov 1 11:33:52 EST 1995


All this sounds very nice, but how about strain differences?
Epicentre indeed sells this buffer "ready-to-go" ( perhaps just suspend 
your fat colony and add DNA...), but my point is that there is a 
two-orders of magnitude difference in how different strains of E.coli
respond to procedures that make them competent. For example, I had
>10E7/ug with the method of Chung on XL-1 Blue, which transforms well
with other protocols, but with SURE we could only exceed this value by
using electroporation... So, beware of simple transformation protocols 
where you need efficiency. With the great number of strains now in use
it is easy to fall into this trap.

Cheers,

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