Molecular Biology and Evolution, Vol 10, 342-361, Copyright © 1993 by Society for Molecular Biology and Evolution
SA Karl and JC Avise
This paper outlines a PCR-based approach for population genetics that
offers several advantages over conventional Southern blotting methods for
revealing restriction-fragment-length polymorphisms (RFLPs) in nuclear DNA.
Primers are constructed from clones isolated from a nuclear DNA library,
and these primers subsequently are employed in in vitro syntheses of
homologous regions. Amplified products are then screened directly for RFLPs
by using gel-staining procedures. Population applications for this
PCR-based approach, including potential strengths and weaknesses, are
exemplified by two RFLP data sets generated to estimate (a) male-mediated
gene flow in the green turtle (Chelonia mydas) and (b) geographic
population genetic structure in the American oyster (Crassostrea
virginica). Restriction assays of amplified products from 14 or 15
independent primer pairs in each species revealed polymorphisms at several
loci that proved highly informative in the population genetic analyses. In
general, the Mendelian polymorphisms produced by this PCR-based approach
will provide useful genetic markers for population studies, particularly in
situations where simpler and less expensive allozyme methods have failed,
for whatever reason, to provide adequate information.
ORIGINAL ARTICLE
PCR-based assays of mendelian polymorphisms from anonymous single-copy nuclear DNA: techniques and applications for population genetics
Department of Genetics, University of Georgia.
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