Goossens B, Waits L P, Taberlet P
Laboratoire de Biologie des Populations d'Altitude (CNRS UMR 5553), Université Joseph Fourier, Grenoble, France.
Mol Ecol. 1998 Sep;7(9):1237-41. doi: 10.1046/j.1365-294x.1998.00407.x.
To test whether plucked hairs are a reliable source of DNA for genotyping microsatellite loci, we carried out experiments using one, three, or 10 hairs per extract for 50 alpine marmots. For each extract, seven independent genotypings were performed for the same locus (multiple-tubes approach). Two types of genotyping errors were recorded: a false homozygote defined as the detection of only one allele of a true heterozygote, and a false allele defined as a PCR-generated allele that was not one of the alleles of the true genotype. Using DNA extracted from one, three, or 10 hairs the overall error rate was 14.00%, 4.86%, and 0.29%, respectively. Based on our results, we conclude that 10 hairs should be used to obtain consistently reliable genotypings using the single-tube approach, and that a single plucked hair could represent a reliable source of DNA if the multiple-tubes approach is used. For future studies of dinucleotide repeat diversity using DNA extracted from one to three shed or plucked hairs, we strongly recommend initiating an appropriate pilot study to quantify the error rate and to determine the reliability of the single-tube approach.
为了测试拔取的毛发是否是用于微卫星基因座基因分型的可靠DNA来源,我们对50只高山旱獭进行了实验,每次提取物分别使用1根、3根或10根毛发。对于每次提取物,对同一基因座进行7次独立的基因分型(多管法)。记录了两种类型的基因分型错误:假纯合子,定义为仅检测到真正杂合子的一个等位基因;假等位基因,定义为PCR产生的、并非真正基因型等位基因之一的等位基因。使用从1根、3根或10根毛发中提取的DNA,总体错误率分别为14.00%、4.86%和0.29%。根据我们的结果,我们得出结论:如果使用单管法,应该使用10根毛发以获得始终可靠的基因分型;如果使用多管法,单根拔取的毛发可能是可靠的DNA来源。对于未来使用从1至3根脱落或拔取毛发中提取的DNA进行二核苷酸重复多样性研究,我们强烈建议开展适当的预实验以量化错误率并确定单管法的可靠性。