Soeing PCR- not getting the mutation- even tho its in the primer

JonnyR9 jonnyr9 at hotmail.com
Fri Oct 12 11:01:40 EST 2001


Dear Beth,

Could you be sequencing the original template?  Whatever contains the
template material survuves until the end of the reaction, perhaps
there's more template than product?

eliza at itsa.ucsf.edu (Beth Holleran) wrote in message news:<4a85aa87.0110091515.47b5a949 at posting.google.com>...
> Hi- I have been doing Soeing PCR for a while but I ran into a new
> problem- this time I get my two products, SOE them together and when I
> sequence them they are Wild type! EVEN THOUGH I used primers that had
> the mutation in them! what in the world has happened? I tried using a
> lot less template (to reduce the amount of WT template) and everything
> has been gel purified.  Has anyone seen this? Help! Please!




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