4种黑曲霉基因组DNA提取方法的比较

Four methods of DNA extraction for Aspergillus niger

目的筛选适合提取曲霉DNA的方法。方法比较2个菌落培养时间段(3 d内和10 d左右)提取DNA质量的差异;运用氯化苄法、石英砂+CTAB法、Biospin法和微波法分别提取黑曲霉基因组DNA,然后用直接电泳、浓度测定、PCR扩增等方法比较所提DNA的浓度和质量。结果培养3 d内的菌落提取的DNA纯度较高,无需纯化即可用于后续实验;4种方法制备的DNA均可用于PCR等后续实验,其中以石英砂+CTAB法提取的DNA纯度好,产率最高。结论石英砂+CTAB法是一种适用于曲霉DNA提取的简便方法。

Objective To find the best DNA extraction method for Aspergillus niger.Methods The purity and output of Aspergillus.niger DNA extracted by benzyl chloride method,CTAB method,Biospin kit and microwave method were compared via agarose gel electrophoresis and PCR.Results The extracted DNA cultured for 3 ds was purer than that for 7-14 ds.All the DNA products were amplifiable for PCR,but the purity and output via CTAB method showed the best results.Conclusions CTAB is suitable for Aspergillus niger DNA extraction.