Sequencing from E.coli.
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.
Klaus Salger phone : +49 (0)89 5902 -502
Zoologisches Institut FAX : -450
AG MacWilliams e-mail: salger at zi.biologie.uni-muenchen.de
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