对中国黄牛毛囊DNA六种提取方法的研究  

Study on Six Methods of Extracting DNA from Hair Follicle of Bos taurina

作　　者：余巨全 巩建华 柯尊伟 Yu Juquan;Gong Jianhua;Ke Zunwei(Shanghai MAP Biotechnology Co.,Ltd.,Shanghai,200090;Hanjiang Normal University,Shiyan,442000)

机构地区：[1]上海迈浦生物科技有限公司,上海200090 [2]汉江师范学院,十堰442000

出　　处：《基因组学与应用生物学》2020年第4期1540-1548,共9页Genomics and Applied Biology

摘　　要：本研究的目的是寻求一种简便、高效和对黄牛没有创伤的DNA提取方法,从而为黄牛基因组DNA的制备提供最优可行方案。本研究以中国黄牛毛囊作为DNA的提取材料,设计了离心柱法、磁珠法、SDS法、CTAB法、碱裂解法和煮沸法等6种DNA提取方法,然后分别对提取的DNA样本进行了分光光度计鉴定、PCR扩增和琼脂糖凝胶电泳。发现在这6种方法中,耗时最短且成本最低的是煮沸法;DNA纯度、浓度和完整度最理想的是CTAB法;而磁珠法可实现大规模高通量提取,能减少实验者的手动工作量,并且适用于基因测序各种常规操作。以此得出,适用于基因测序的6种黄牛毛囊DNA提取方法中最优的是磁珠法。本研究对中国黄牛毛囊DNA的6种提取方法进行了总结和分析,为今后提取毛囊DNA的研究提供了借鉴和参考。This study aimed to find a simple,efficient and non-invasive DNA extraction method which might be the best solution for the preparation of genomic DNA of yellow cattle.We used hair follicles of Bos taurinaas materials to extract DNA with six DNA extraction methods including spin column method,magnetic bead method,SDS method,CTAB method,alkaline lysis method and boiling method.We then analyzed these six groups of hair follicles DNA samples with spectrophotometer identification,PCR amplification and agarose gel electrophoresis.The result showed that among the six methods,boiling method is the least time-consuming and lowest cost.The CTAB method performed the best purity,concentration and integrity of DNA extracted.The magnetic bead method can achieve large-scale high-throughput extraction of DNA and reduce the amount of manual labor.The magnetic bead method is suitable for various routine operations of the gene sequencing.The comprehensive analysis of six methods showed that the best method to extract DNA from Hair Follicle of Bos taurinais magnetic bead method in gene sequencing.In this paper,six methods of DNA extraction from Hair Follicle of Bos taurinais were summarized and analyzed,which provided reference for future research on DNA extraction from cattle hair follicles.

关 键 词：牛毛囊DNA DNA提取 PCR扩增 

分 类 号：S823.81[农业科学—畜牧学]