Oligo synthesis woes

Dr. C. Dolphin cdolphin at hgmp.mrc.ac.uk
Fri Aug 26 07:48:17 EST 1994


I have recently synthesised 6 oligos on an ABI DNA synthesiser
directly after the machine was serviced and fitted with fresh
chemicals. The first and last oligos were synthesised on ABI
columns while for the middle four I used Prime Synthesis columns
designed for the ABI system. Yields for all 6 were OK but only the
primers made on the ABI columns work as primers in PCR -  the
other four don't work at all in PCRs (all are tested with opposite
primers that I know are fine). Has anyone ever experienced anything
like this ? My best guess at the moment is that while synthesis is
OK (hence the yield) maybe there's a problem during removal of the
oligo from the column that produces an altered 3' base that is unable
to accept dNTPs in the PCR. Anyone got any ideas?

Colin



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