Primer synthesis problems
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.
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
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