一种简便高效的酵母基因组提取方法

提取基因组进行检测是酵母研究过程中的必要步骤之一。以毕赤酵母菌株GS115作为研究对象,主要成分为0.2 mol/L醋酸锂和1% SDS的酵母裂解液能高效的裂解酵母细胞壁。与两种酵母基因组提取试剂盒相比,该方法从相同体积的酵母培养液中获得的基因组的量高5倍以上,并且操作简便、快速,能在2 h内完成一次提取过程,极大地缩短了时间。以GS115中的内源AOX基因为目的基因,对提取的基因组进行PCR检测和Southern杂交检测,进一步验证了基因组的质量。因此,本文建立了一种简便、快速、经济而高效的酵母基因提取方法。

A Simple and Effective Method for Genomic DNA Extraction from Yeasts

Extraction genomic DNA from yeasts for some assays is very important. Yeast-lysis solution contained 0.2 mol/L lithium acetate, 1% SDS and other components can efficiently lysis Pichia pastoris GS115 cells. In comparison with two kinds of yeast genomic DNA extraction kits, the amount of genomic DNA extracted by this way was more than five times as much as those by the kits from same volume liquid culture. Moreover, the genomic DNA extraction protocol set up in the paper was simple and easy to perform. It took 2 hours to finish a whole protocol. To further test the quality of genomic DNA, PCR amplification and Southern blot analysis of AOX gene in yeast genomic DNA were done. Therefore, we have developed a quick, low cost and effective genomic DNA extraction protocol from yeast cells in this paper.