Competitive blocker pcr
Kimberly Lieuallen
lieuallen1 at LLNL.GOV
Mon Nov 11 15:42:05 EST 1996
Hello everyone,
We are interested in utilizing competitive blocker pcr to screen
bacterial colonies for the presence or absence of a transposon in
the insert. I have read an abstract entitled: Allele-specific
competitive blocker PCR: a one-step method with applicability to pool
screening by Orou, A, et al. In this procedure three primers are used
in the same pcr reaction but one primer acts as a blocker to
amplification since the 3' end of the primer is labelled with a
dideoxynucleotide. In their paper, the "crippled" primer successfully
blocked the wildtype allele, so that only detection of the mutant
alleles resulted.
Has anyone has experience with this method? Is the amplification of
the wildtype allele practically undetectable?
If anyone has experience success/disgust with this method please email
me at: lieuallen1 at llnl.gov
Thanks,
Kim
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