摘要
收集九龙山自然保护区野外自然条件下的黑麂粪便样品,用无水乙醇保存于-70℃冰箱,经十六烷基三甲基溴化铵(CTAB)裂解液温浴粪便、酚/氯仿抽提及聚合酶链反应(PCR)纯化试剂盒纯化等步骤提取黑麂粪便DNA,并扩增线粒体DNA细胞色素b基因和2个微卫星位点进行检测;同时提取黑麂肌肉和皮毛样品的DNA作为对照.结果显示,该方法可以从粪便中提取到高质量的黑麂DNA,并可进行有效扩增,为黑麂及其他濒危鹿科动物的非损伤取样提供了一种简易方法.
The feces of Muntiacus crinifrons were collected from field in Jiulong Mountain Nature Reserve, preserved in absolute ethanol, and stored in -70℃ refrigeratory. The DNA in feces was extracted by procedures including digestion in CTAB buffer, extraction by phenoL/chloroform and purification by PCR Cleanup Kit, and then amplified by primers of mitochondrial DNA cytochrome b gene and two microsatellite loci for test. As positive control, the DNA of muscle and skins of Muntiacus crinifrons were extracted and amplified respectively. The results showed that this method could produce high quality DNA from feces, and the product could be effectively amplified by the primers. The method gave facilities for noninvasive sampling of Muntiacus crinifrons and other endangered deers.
出处
《浙江师范大学学报（自然科学版）》
CAS
2008年第2期214-219,共6页
Journal of Zhejiang Normal University:Natural Sciences
基金
国家自然科学基金资助项目(30370219)
