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- Title
不同保存方法对川金丝猴粪便DNA提取效果的影响.
- Authors
周芸芸; 张宇; 杨万吉; 蒋军; 余辉亮; 李迪强; 张于光
- Abstract
Objective: Sichuan snub-nosed monkey (Rhinopithecus roxellana), a endemic rare species in China and to be list to a national first class protected animal species, has characteristic of natural alert and arboreal life. Fecal, as a non-invasive sample, gives great convenience to researches on population survey, genetic diversity, genetic relationship, phylogeny. However, appropriate sampling and preservation methods are the premise to get useful DNA for the researches of endangered animals with non-invasive fecal sample. This study attempts to establish a high efficient, convenient method. Methods: Combined the reported preservation methods of the endangered animal fecal samples with the actual fact, three methods included drying, freezing and drying-freezing are used. Taking blood samples and hair samples as a reference, it is to analyze the effects of different fecal samples preservation methods on the efficiency of DNA extraction and the success rate of mtDNA and microsatellite DNA loci gene amplification, which included drying, freezing and drying-freezing in 1 week, 2 months and 6 months storage time respectively. Results: The amplification success rate of mtDNA gene is 92% with the three preservation methods and microsatellite loci amplification success rate of drying, freezing and drying-freezing was 79%, 78%, 80% respectively after a week. For the mtDNA gene amplification success rate with drying method, freezing method and drying-freezing method, it is 80%, 76%, 80% respectively after two months, and 56%, 52% and 64% respectively after six months. For microsatellite loci success rate with three methods, it is 65%, 61% and 67% respectively after two months, 40%, 34% and 46% respectively after six months. It showed that DNA amplification success decreases with the increasing of the storage time. Compared with two other methods, the samples with drying-freezing method can get better DNA extracted quality and high amplification success rate of mtDNA gene and microsatellite loci. Conclusion: The fecal of Sichuan snub-nosed monkey can be the research sample for providing effective information on some genetics study such as the evaluation of genetic diversity. Compared with other two methods, the drying-freezing method for fecal preservation with long-way transportation is a better way to get quality DNA extracted and high efficiency of gene amplification.
- Publication
Progress in Modern Biomedicine, 2018, Vol 18, Issue 1, p43
- ISSN
1673-6273
- Publication type
Article
- DOI
10.13241/j.cnki.pmb.2018.01.009