: > I've got a problem with a PCR that I'm attempting...
: > I'm trying to use a pair of primers to follow a series
: > of deletions that I'm making in a plasmid.
: > Under 'standard' conditions I can amplify the deleted
: > version of the plasmid but not the original undeleted
: > version.
: > Can anyone explain this, I've tried all the usual
: > variations in the reaction conditions, etc. with no luck!
Is your plasmid ( the undeleted one) old? Apparently this happens if the
DNA is old or sits around too long. I don't know why but maybe some
modifications of Primer binding site happens in the intact one while not
in the deleted versions because they are worked on quickly.
Program in Molecular Biology
Dept.of Chemistry and Biochemistry Box3C
NMSU Las Cruces NM