Sequencing from E.coli.

Klaus Salger salger at wap1.zi.biologie.uni-muenchen.de
Wed Nov 15 18:05:37 EST 1995


Paul Bundockj (bundock at rulsfb.leidenuniv.nl) wrote:
: Hi there, I'm trying to sequence ds plasmid DNA  rescued in the E.coli
: strain MC1061.So far its been very difficult to sequence mini prep DNA 
: although it should be possible if the prep is reasonably clean.This 
: problem with the E.coli background seems common, does anybody have a 
: list or reference for E.coli backgrounds and their suitability for 
: sequencing. By the way,I'm using T7 polymerase and 1.5-2 micro grams 
: template. Thanks a lot, Paul Bundock.

MC1061 doesn't carry the endA mutation. It seems to be difficult to get rid
of this endonuclease which makes sequencing very difficult. Switching to
an endA- strain (DH5a, XL1,...) should fix this problem.

Cheers
  Klaus

--
Klaus Salger                phone : +49 (0)89 5902 -502
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