How to use Expand PCR System
vnp at mole.bio.cam.ac.uk
Tue Sep 17 06:19:44 EST 1996
In article <323E6ED2.603B at bsa.bristol.ac.uk>, "Andrew J. Doherty"
<doherty at bsa.bristol.ac.uk> wrote:
> ouyhs at PUBLIC.CC.JL.CN wrote:
> > Dear Netter:
> > I have bought a Expand Long Template PCR System and a Expand High
> > Fidelity PCR System From Boehringer Mannheim. I wish I could use
> > these kits to amplify and clone a gene about 10kb from bovine genome
> > DNA. But I do not know which kind end be produced in the PCR products
> > if we use those Expand systems. Can I use the T-vctor to clone the
> > PCR products? Do you have any literature on amplifing and cloning
> > the DNA by using those Expand systems?
> The data sheets for the Expand High Fidelity say that the enzyme
> produces a mixture of blunt and T-overhang ends, like Taq. So youshould
> be able to use T/A cloning. I haven't personally done T/A cloning with
> this enzyme(s); we use it for it's proof reading in mutagenesis and cut
> the ends of the PCR products.
yes, you can use T/A cloning. i've used kits from both invitrogen and promega
with success. good luck.
Venkat Pisupati Lab -44-01223-334131
Wellcome/CRC Institute Fax -44-01223-334134
Dept. of Genetics E.mail. vnp at mole.bio.cam.ac.uk
University of Cambridge
UK CB2 1QR
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