Dear ABRF'ers:
Theoretically, I would think that a primer used for PCR should double as a
sequencing primer, provided one has cleaned the PCR product completely from the
two primers. However, frequently it does not. One of our users has some BACs
from which he's PCRed up a fragment. After gel purification and extraction from
the gel, he's tried to use the PCR primers for sequencing and has generally
failed miserably. No sequence at all (though there is a "T" blob at ~110 bp -
now what is the cause of that?). Any general procedures - or references thereto
- for sequencing PCR products will be appreciated. Thanks,
Mark J. Miller, Ph.D.
Manager: DNA Sequencing MiniCore Facility
Bldg 37, Rm 3C28, MSC 4255
DBS/NCI/NIH
Bethesda, MD 20817-4255
301-496-5688 x226
mark_james_miller@nih.gov
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