晋西北酸粥发酵过程中微生物基因组DNA提取方法的比较
Comparison of microbial genome DNA extraction methods used in the fermentation process of northwestern Shanxi acid gruel
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摘要: 为了找到一种简单高效的晋西北酸粥发酵过程中微生物基因组DNA提取方法,本文采取对提取的微生物基因组DNA进行浓度测定的方法,比较了溶菌酶-SDS-蛋白酶K法、改良CTAB法和改良CTAB-溶菌酶法三种方法的提取效果。结果表明:溶菌酶-SDS-蛋白酶K法效果最好,可以获得质量较高的微生物基因组DNA,DNA浓度为1657.53 ng/μL;改良CTAB-溶菌酶法比溶菌酶-SDS-蛋白酶K法效果差,DNA浓度为1308.08 ng/μL;改良CTAB法对微生物基因组DNA的提取效果比溶菌酶-SDS-蛋白酶K法更差,DNA浓度为1098.36 ng/μL。以提取到的微生物基因组总DNA为模板,进行16S r DNA基因的PCR扩增,在回收产物中目的条带清晰,表明提取到的微生物基因组DNA含量高、结构完整。整体实验结果表明,溶菌酶-SDS-蛋白酶K法对于提取晋西北酸粥发酵过程中微生物基因组DNA效果最好。Abstract: In order to obtain a simple and efficient extraction method of microbial genome DNA from northwestern Shanxi acid gruel,three extraction methods(lysozyme-SDS-proteinase K method,improved CTAB method and improved CTAB-lysozyme method) were compared according to the concentrations of extracted microbial genome DNA. The results showed that the effect of the lysozyme-SDS-proteinase K method was the best,flowed by improved CTAB-lysozyme method and improved CTAB method. By lysozyme-SDS-proteinase K method,higher quality of microbial DNA could be obtained and the DNA concentration was 1657.53 ng/μL.When using the improved CTAB-lysozyme method and improved CTAB method,the DNA concentrations were respectively 1308.08 ng/μL and 1098.36 ng/μL. The total DNA of 16 S r DNA gene PCR amplification was tested.The results showed that the purpose stripe was clear in recycling products,which indicated that the microbial genome DNA obtained in this experiment had high content and complete structure. All the results showed that the lysozyme-SDS-proteinase K method was the best to extract the microbial genome DNA of northwestern Shanxi acid gruel.
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