Karl S A, Avise J C
Department of Genetics, University of Georgia.
Mol Biol Evol. 1993 Mar;10(2):342-61. doi: 10.1093/oxfordjournals.molbev.a040002.
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.
本文概述了一种基于聚合酶链反应（PCR）的群体遗传学方法，与传统的Southern印迹法相比，该方法在揭示核DNA中的限制性片段长度多态性（RFLP）方面具有多个优势。引物由从核DNA文库中分离出的克隆构建而成，随后这些引物用于同源区域的体外合成。然后通过凝胶染色程序直接筛选扩增产物中的RFLP。基于PCR的这种方法在群体中的应用，包括潜在的优点和缺点，通过两个RFLP数据集得到例证，这两个数据集用于估计：（a）绿海龟（蠵龟）中雄性介导的基因流动，以及（b）美国牡蛎（弗吉尼亚牡蛎）的地理群体遗传结构。对每个物种中14或15个独立引物对的扩增产物进行限制性分析，发现在几个位点存在多态性，这些多态性在群体遗传分析中被证明具有很高的信息量。一般来说，这种基于PCR的方法产生的孟德尔多态性将为群体研究提供有用的遗传标记，特别是在由于任何原因，更简单、成本更低的等位酶方法未能提供足够信息的情况下。
