Sequencing vector needed
gcantor at VETMED.WSU.EDU
Sat Oct 15 19:28:37 EST 1994
> On 13 Oct 1994, f.r.byrne wrote:
> > --
> > I need some help with a sequencing problem.
> > I want to obtain a vector for sequencing a PCR
> > product which is preferably high copy number
> > and does not have ampicillin or tetracycline
> > resistance as a marker.
> > it is for use in E. coli .
> > it should have standard cloning sites, specifically
> > EcoRI.
> > I have tried the ususal source sand contacted
> > pretty much every comopany but no luck.
> > if anybody can help then please post a message
> > or email me at f.r.byrne at abdn.ac.uk
> > Any help gratefully received.
> > thanks.
> > Fergus Byrne
> > Aberdeen University, Scotland.
> Why not sequence the PCR product directly? It is much quicker than
> cloning into a vector and then sequencing, if your goal is to obtain a
> sequence. U.S. Biochemical has a kit for doing this, and perhaps other
> companies do as well. It is based on amplifying your PCR product with
> primers and then digesting the primers and NTP's with Exonuclease I and
> with shrimp Alkaline Phosphatase, then deactivating the shrimp Alk Phos
> by heating the sample.
> Glenn Cantor
> Department of Veterinary Microbiology and Pathology
> Washington State University
> Pullman WA 99164-7040
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