Cleanup of short PCR Product

LeeSylvers leesylvers at aol.com
Sat Jun 29 14:02:44 EST 1996


An alternate way to prepare PCR products for sequencing is to have one of
the primers used in the PCR biotinylated.  The PCR product can then be
captured on a solid support (ideally magnetic due to ease of use and less
lose of sample).  Once this is done, the non-biotinylated strand can be
released and used for sequencing.  In this case, not only have you
eliminated any primer contamination but also now have a single-stranded
product to sequence, which should yield longer and cleaner sequence reads.
 It is also possible to do solid-phase sequencing on the immobilized
strand.
CPG, Inc. has a complete kit which allows you to carry this prorocol out
with ease.  It includes the magnetic particles, all necessary buffers, and
universal primers (biotinylated and not biotinylated) that flank most
multiple cloning sites.



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