Hind III Digest of PCR Products

Aawara Chowdhury aawara at nohome.org
Sun Jan 29 14:54:03 EST 2006


In <mailman.891.1138561310.29584.methods at net.bio.net>,
 Mark Blenner <markblenner at yahoo.com> wrote:

> I have a 790 bp PCR fragment I am trying to ligate
> into a 7400bp vector - some data suggests there may be
> some difficulties in cutting the HindIII  at the end
> of my PCR fragment, but I've added 6 extra basepairs
> and still doesnt work. Anyone seen this before? 
>

Usually not - what's the base composition of the terminal 6 bp?  In any event,
here's one solution.  Phosphoryate the oligos before PCR.  Purify the PCR 
product, and self-ligate it.  Now cut this with HindIII.

AC
-- 
In America, through pressure of conformity, there is freedom of choice,
but nothing to choose from - Peter Ustinov.


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