文档介绍：
秦岭川金丝猴及伴生植食兽类粪便基因组
DNA 提取方法研究#
陈丹1,黄康1,胡永乐2,张红阳1,李保国1,郭松涛1**
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(1. 西北大学生命科学学院,西安 710069;
2. 陕西周至国家级自然保护区管理局,西安 710400)
摘要:粪便作为濒危野生动物的遗传学样品日益成为动物保护研究的重要材料,如何保存在
野外获得的粪便样品并从其中提取高质量的 DNA 成为此类研究的难点。不同物种的粪便在
不同气候环境的保存时效各有差异。本研究主要以秦岭地区川金丝猴及伴生兽类的粪便样品
作为研究材料,选用随机微卫星位点以及一对通用线粒体 D-Loop 区域的引物,对提取的
DNA 进行扩增,以检测所提取 DNA 样品的质量。优化常用粪便样品 DNA 的提取方法后,
通过对野外放置不同时间的样品所提取的 DNA 的质量进行比较,确定野外条件下粪便样本
的有效采集期。结果显示秦岭地区野生金丝猴及伴生植食兽类粪便样品在野外露天保存时间
较长的冬季,金丝猴粪便在 3 天内可以提取有效采的基因组 DNA 用于微卫星分型,线粒体
扩增等下游实验。优化保存和提取方法同样适用于羚牛、斑羚、大熊猫、小熊猫的研究。
关键词:动物学;川金丝猴;非损伤采样;粪便;微卫星;线粒体
中图分类号:Q95
A new protocol for DNA extraction of faece samples from
QinLing snub-nosed monkey and herbivorous mammals
CHEN Dan1, HUANG Kang1, HU Yongle2, Zhang Hongyang1, LI Baoguo1,
GUO Songtao1
(1. College of Life Sciences, Northwest University, Xi'an 710069;
2. Administration of ZhouZhi National Nature Reserve, Xi'an 710400)
Abstract: In recent years, faeces samples have been extensively used in ic studies of
endangered wild species. However, How to preserve stool sample, and extract high quality DNA
e difficulties during the experimental procedures. In this article, DNA were extracted from
faece samples of sichuan snub-nosed monkey and were amplified in 5 randomly selected
polymorphic SSR loci and one pair of 646bp mitochondrial D-Loop gene in order to text the
quality of DNA extract. By referencing the reported preservation and extraction methods, we have
developed an efficient and economic method to extract DNA from these faeces samples. By
comparing the DNA quality from different period after excretion ,the effective collecting time
span were determined where faece samples collected in this interval were able to provide high
quality template for downstream molecular biology experiments. q By applying this protocol to
some other herbivorous mammals from Qinling mountains The results show, we obtained it was
reliable and valid in herbivorous species.
Keywords: Zoology; Sich