Quantitative polymerase chain reaction analysis of DNA from noninvasive samples for accurate microsatellite genotyping of wild chimpanzees (Pan troglodytes verus)

被引:362
|
作者
Morin, PA [1 ]
Chambers, KE [1 ]
Boesch, C [1 ]
Vigilant, L [1 ]
机构
[1] Max Planck Inst Evolutionary Anthropol, D-04103 Leipzig, Germany
关键词
chimpanzees; faeces; microsatellite genotyping; noninvasive samples; paternity; quantitative PCR;
D O I
10.1046/j.0962-1083.2001.01308.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Noninvasive samples are useful for molecular genetic analyses of wild animal populations. However, the low DNA content of such samples makes DNA amplification difficult, and there is the potential for erroneous results when one of two alleles at heterozygous microsatellite loci fails to be amplified. In this study we describe an assay designed to measure the amount of amplifiable nuclear DNA in low DNA concentration extracts from noninvasive samples. We describe the range of DNA amounts obtained from chimpanzee faeces and shed hair samples and formulate a new efficient approach for accurate microsatellite genotyping. Prescreening of extracts for DNA quantity is recommended for sorting of samples for likely success and reliability. Repetition of results remains extensive for analysis of microsatellite amplifications beginning from low starting amounts of DNA, but is reduced for those with higher DNA content.
引用
收藏
页码:1835 / 1844
页数:10
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