Multiplex preamplification PCR and microsatellite validation allows accurate single nucleotide polymorphism (SNP) genotyping of historical fish scales

Incorporating historical tissues into the study of ecological, conservation, and management questions can broaden the scope of population genetic research by enhancing our understanding of evolutionary processes and anthropogenic influences on natural populations. Genotyping historical and low-quality samples has been plagued by challenges associated with low amounts of template DNA and the potential for preexisting DNA contamination among samples. We describe a two-step process designed to (i) accurately genotype large numbers of historical low-quality scale samples in a high-throughput format and (ii) screen samples for preexisting DNA contamination. First, we describe how an efficient multiplex preamplification PCR of 45 single nucleotide polymorphisms (SNPs) can generate highly accurate genotypes with low failure and error rates in subsequent SNP genotyping reactions of individual historical scales from sockeye salmon (Oncorhynchus nerka). Second, we demonstrate how the method can ...

将历史生物组织样本纳入生态学、保护生物学与资源管理相关研究，可通过深化对自然种群进化过程及人为活动影响的认知，拓宽种群遗传学（population genetics）研究的范畴。对历史样本与低质量样本开展基因分型（genotyping）工作，长期以来饱受模板DNA含量极低、样本间潜在预先存在DNA污染等技术难题的困扰。本研究介绍一种双流程实验方案，旨在实现两大目标：（i）以高通量（high-throughput）方式精准对大量历史低质量鱼鳞样本进行基因分型；（ii）筛查样本是否存在预先存在的DNA污染。首先，本研究详述了针对45个单核苷酸多态性（Single Nucleotide Polymorphisms, SNPs）位点的高效多重预扩增聚合酶链式反应（Polymerase Chain Reaction, PCR），如何在后续针对红大麻哈鱼（Oncorhynchus nerka）单条历史鱼鳞样本的SNP基因分型反应中，获得低失败率与低错误率的高精度基因型数据。其次，本研究证实了该方法可用于……