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野生黑猩猩幼崽的尿液和唾液可通过非侵入性方式采集用于DNA分析。

Wild chimpanzee infant urine and saliva sampled noninvasively usable for DNA analyses.

作者信息

Inoue Eiji, Inoue-Murayama Miho, Takenaka Osamu, Nishida Toshisada

机构信息

Human Evolution Studies, Department of Zoology, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan.

出版信息

Primates. 2007 Apr;48(2):156-9. doi: 10.1007/s10329-006-0017-y. Epub 2006 Nov 17.

Abstract

In many genetic studies on the great apes, fecal or hair samples have been used as sources of DNA. However, feces and hairs are difficult to collect from chimpanzee infants under 3 years of age. As alternative DNA sources, we investigated the efficiency of collecting urine samples from infants compared with fecal samples, as well as the validity of the DNA extracted from urine and saliva samples of well-habituated M group chimpanzees (Pan troglodytes schweinfurthii) in the Mahale Mountains National Park, Tanzania. We collected 40 urine and 3 fecal samples from 10 infants under 3 years. Compared with feces, the urine samples were relatively easy to collect. The saliva of infants, which remained on the twigs sucked by them, was collected using cotton swabs. The average amounts of DNA extracted from the 40 urine and 6 saliva samples were 3,920 and 458 pg/mul, respectively. The rate of positive PCR was low and the allelic dropout rate was high when using less than 25 pg of template DNA in the PCR mixtures. Based on the amounts of DNA, 50% of the urine samples and 100% of the saliva samples were judged usable for accurate microsatellite genotyping. For infant chimpanzees in particular, collecting urine and saliva as an alternative to fecal and hair samples can reduce the effort invested in collection in the field.

摘要

在许多关于大猩猩的基因研究中,粪便或毛发样本一直被用作DNA来源。然而,对于3岁以下的黑猩猩幼崽来说,粪便和毛发很难采集。作为替代的DNA来源,我们调查了从幼崽采集尿液样本与粪便样本相比的效率,以及从坦桑尼亚马哈尔山国家公园习惯良好的M群黑猩猩(Pan troglodytes schweinfurthii)的尿液和唾液样本中提取的DNA的有效性。我们从10只3岁以下的幼崽中采集了40份尿液样本和3份粪便样本。与粪便相比,尿液样本相对容易采集。用棉签采集了幼崽留在它们吸吮过的树枝上的唾液。从40份尿液样本和6份唾液样本中提取的DNA平均量分别为3920和458 pg/μl。当PCR混合物中使用少于25 pg的模板DNA时,PCR阳性率较低,等位基因缺失率较高。根据DNA量,50%的尿液样本和100%的唾液样本被判定可用于准确的微卫星基因分型。特别是对于黑猩猩幼崽,采集尿液和唾液作为粪便和毛发样本的替代方法可以减少在野外采集时投入的精力。

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