tjstbc at sd.cts.com
Thu Feb 8 13:46:52 EST 1996
You should suspect the oligos used for PCR - many oligos that are
synthesized have mutations due to the chemistry used for oligo
synthesis. The best way to check for restriction sites in the PCR
product is to subclone the PCR fragment into a TA cloning vector and
sequence through the ends to check for the proper restriction sites.
You may want to PAGE purify the oligos before using them for PCR - the
PCR fragments resulting from PAGE purified oligos (as opposed to OPC
purified) seem to be more "cloneable".
More information about the Methods