一种简易有效的提取真菌DNA化学改良方法

A Simple and Effective Chemical Modification Method for Extracting DNA from Fungi

[目的]探索适合真菌基因组DNA提取的有效方法。[方法]以酵母菌(Hansenulasp.HS07A)和青霉菌(Penicilliumsp.HS07B)为供试菌株,分别采用煮沸裂解法、石英砂研磨法、研磨和反复研磨冻融法和化学改良法提取其基因组DNA,比较这4种方法的提取效果。[结果]化学改良法提取基因组DNA,除去了机械破壁过程,减少了机械力对DNA造成的破坏,运用阴离子去污剂SDS提取DNA安全、价廉,能有效、完整地提取高质量的基因组DNA,凝胶电泳条带清晰、无弥散,以此基因组DNA作为目的片段的PCR扩增模板,扩增效果优于改良前的方法,并且这种方法也适合多种真菌和细菌基因组DNA的提取。[结论]化学改良法是提取真菌基因组DNA的有效方法。

[ Objective] The aim was to explore the effective method suitable for the extraction of fungal genomic DNA. [ Method] With Hansenu/a sp. HSOTA and Penicillium sp. HST/B as tested strains,boiling lysis method,quartz sand grinding method,grinding and repeated grinding and freeze-thaw method,and chemical modification method were adopted for the extraction of their genomic DNA and the extraction effect of the 4 methods were compared. [ Resuh] Using chemical modification method extracting genomic DNA could eliminate the process of machine wall-breaking,reduce the damage of mechanical force on DNA. Using anionic detergent SDS for DNA extraction was safe and cheap,which could extract high quality genomic DNA effectively and completely. The gel electrophoretic band was clear and had no dispersion,and using the genomic DNA as the tearplate for PCR anplification of target segment, the amplification effect was better than that by the method before modification and the method was still suitable for the genomic DNA extraction of many kinds of epiphyte and bacteria. [ Conclusion] Chemical modification method was an effective method for fungal genomic DNA extraction.