藏酋猴粪便总DNA提取效果的比较研究

Comparative Study on the Extraction Effect of Total Fecal DNA of Macaca thibetana

近年来,野生动物的保护研究已成为一大热点,其采样方法主要以非损伤性取样(粪便)为主。为筛选一种操作简单、经济成本较适宜的动物粪便基因组DNA高效提取试剂盒,笔者选用QIAamp Fast DNA Stool Mini Kit(德国:粪便DNA纯化试剂盒)、生工磁珠法基因组DNA抽提试剂盒、Stool Genomic DNA Kit(康为世纪:粪便基因组提取试剂盒)、生工Ezup柱式土壤DNA抽提试剂盒这4种来自不同厂家,其使用频率较高及市售商业化的试剂盒,各自提取4份新鲜藏酋猴(Macaca thibetana)粪便样本的基因组DNA。通过比较各试剂盒所提取基因组DNA的浓度、纯度,并对其16S rRNA进行PCR扩增及singer测序,从而获得不同试剂盒基因组DNA的提取效果。比较发现,Stool Genomic DNA Kit(康为世纪)的效果较好且经济,并将其应用于兽类(野猪),灵长类(黔金丝猴),鸟类(黑颈鹤、灰鹤、斑头雁和红腹角雉)等物种进行验证,发现均获得理想的基因组DNA提取效果,说明Stool Genomic DNA Kit(康为世纪)可作为鸟兽等野生动物粪便基因组DNA提取的有效试剂盒。这一研究结果为其他类似研究提供了参考依据。

In recent years,the research on wildlife protection has become a hot spot,and its sampling method is mainly non-invasive sampling(feces).In order to screen an efficient extraction kit of animal fecal genomic DNA with simple operation and appropriate economic cost,this study mainly selects QIAamp fast DNA stool Mini Kit(Germany:fecal DNA Purification Kit),Mag-MK Animal Genomic DNA Extraction Kit,Stool Genomic DNA Kit and Ezup Column Soil DNA Purification Kit from different manufacturers,The kit,which is frequently used and commercially available,extracts genomic DNA from four fresh fecal samples of Macaca thibetana.By comparing the concentration and purity of genomic DNA extracted by each kit,and PCR amplification and singer sequencing of 16S rDNA,the extraction effects of genomic DNA from different kits were obtained.It is found that the effect of Stool Genomic DNA Kit is good and cost-effective.It is applied to animals(Sus scrofa),primates(Rhinopithecus brelichi),birds(Grus nigricollis,Grus grus,Anser indicus and Tragopan temminckii)and other species for verification.It is found that ideal genomic DNA extraction effects are obtained,It shows that Stool Genomic DNA Kit can be used as an effective kit for extracting genomic DNA from feces of wild animals such as birds and animals.This research result provides a strong reference basis for other similar studies.