Primer synthesis problems

Paul Shinn pshinn at force.stwing.upenn.edu
Mon Dec 6 11:54:30 EST 2004


Duncan Clark <blackhole at abuse.plus.com> wrote:
:>
:>Try TA cloning a few PCR products and sequence just the ends.
: 
: To be clearer, I mean TA clone before any digest. Raw PCR products.
: 
: Duncan

About 6 months ago when I was in another synthesis slump,
I did this and I found both ends of a clone to be good 35%
of the time.  This makes sense to me since (theoretically)
my forward 56mer should have 56% full length ends and my
43mer 64%.  Of course things change in  6 months and I am
planning on trying this again.


To the other poster, I would love to cartridge purify these
oligos but I am doing hundreds and it is cost prohibitive.

We do an overnight digest with the Sfi because we've found that
anything less than 8 hours is insufficient and gives us poor
transformants.

Thanks again, Paul



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