用Chelex-100法大规模快速提取牦牛肌肉中基因组DNA

Rapid and large-scale extraction of genomic DNA from yak muscles using Chelex-100 method

旨在建立快速、大规模提取牦牛(Bos grunniens)肌肉中基因组DNA的方法.试验样品为冰冻保存的九龙牦牛背最长肌(n=10).采用两种取样方法(镊子取样和枪头取样)采集冰冻的肌肉样品,用Chelex-100方法提取其中的基因组DNA,并进行PCR扩增.结果显示,镊子取样和枪头取样提取的基因组DNA,均可用于两个核基因和一个线粒体基因的PCR扩增,但枪头取样简便易行,提取的DNA用PCR扩增细胞色素b,扩增35个循环数的PCR产物可直接测序.表明Chelex-100法可用于快速提取牦牛肌肉中基因组DNA,操作简单,成本低廉,适用于大规模提取组织中的DNA.

The objective of this study was to establish a rapid and large-scale method for the extraction of genomic DNA from yak （Bos grunniens） muscles. Ten frozen longisimus dorsi samples of Jiuloug yaks were tested. Two sampling methods （ tweezers sampling and pipette tips sampling） were used for the sampling of frozen muscles. The genomic DNA was further extracted with the Chelex-100 method followed by PCR amplification. The results showed that the genomic DNA extracted with both sampling methods could be used for the PCR amplification of two nuclear genes and one mitochondrial gene. Pipette tips sampling was simple and easy, and the PCR products of cytochrome b gene with 35 cycles could be sequenced directly. The present result showed that the Chelex-lO0 method can be used for rapid extraction of genomic DNA from yak muscles, exhibiting simple and inexpensive advantages, and is suitable for large-scale extraction of DNA from tissues.