Sequencing vector needed

Glenn Cantor 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
> USA 
> 
> 



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