PCR sequencing directly from E.coli colonies
logan at msdos.montpellier.inra.fr
Tue Apr 11 05:12:23 EST 1995
I've been using a cycle sequencing kit for a while (Pharmacia's) and have always
prepared my DNA before carrying out the reactions using either a miniprep
protocol or "Wizards". As I should like to run a gel before the end of the week,
and I've only just carried out my deletions, I was very interested in the
protocol that Pharmacia detail, where a colony is lysed in TAPS buffer/boiled
and the supernatant used in the sequencing reactions. This sounds perfect, but I
don't want to waste a gel if it gives lousy results, false economy of time! So
if anyone out there has used this and can comment I'd appreciate the advise.
Also, bit ignorant maybe, but I've never heard of TAPS, can anyone enlighten me?
Thanks in advance
Logan at montpellier.inra.fr
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