Multiplex preamplification PCR and microsatellite validation enables accurate single nucleotide polymorphism genotyping of historical fish scales.

摘要: Incorporating historical tissues into the study of ecological, conservation and management questions can broaden scope population genetic research by enhancing our understanding evolutionary processes anthropogenic influences on natural populations. Genotyping low-quality samples has been plagued challenges associated with low amounts template DNA potential for pre-existing contamination among samples. We describe a two-step process designed to (i) accurately genotype large numbers scale in high-throughput format (ii) screen contamination. First, we how an efficient multiplex preamplification PCR 45 single nucleotide polymorphisms (SNPs) generate highly accurate genotypes failure error rates subsequent SNP genotyping reactions individual scales from sockeye salmon (Oncorhynchus nerka). Second, demonstrate method be modified amplification microsatellite loci detect A total 760 182 contemporary fin clip were genotyped screened exceedingly similar both Pre-existing 21% was successfully identified screening amplified loci. The advantages automation, rates, ability combine make protocol ideally suited efficiently potentially contaminated sources DNA.