Primer synthesis problems

Duncan Clark blackhole at abuse.plus.com
Mon Dec 6 04:31:46 EST 2004


Historians believe that in newspost <cor4t6$dp7n$1 at netnews.upenn.edu> on 
Sat, 4 Dec 2004, Paul Shinn <pshinn at force.stwing.upenn.edu> penned the 
following literary masterpiece:
>Can anyone address these issues?  The synthesizer is doing the best it
>can do.  I don't see any problems with it but am I missing something?
>Could our cloning problems be from bad vector?

Try TA cloning a few PCR products and sequence just the ends. If the Sfi 
I sites are all intact then the problem is the Sfi I digest and /or 
ligation. If the sites are intact change your enzyme source/batch/PCR 
clean up solutions etc etc.

If the sites are wrong then it is a DNA synthesiser fault or say bad or 
even mixed phosphoramidite stocks.

If the sites are missing then bad synthesis.

Duncan
-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.



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