腐乳中菌群基因组DNA提取方法的对比研究

Comparison of methods of genomic DNA extraction from microorganism in sufu

为更好研究腐乳中微生物菌群的多样性,采用改良十六烷基三甲基溴化铵(CTAB)法和玻璃珠法对腐乳中的微生物进行非培养方式的基因组DNA提取,并将提取产物进行高通量测序分析。结果显示,在基因组DNA提取浓度方面,玻璃珠法和改良CTAB法均能得到一定量的基因组DNA,且均可得到扩增条带,但玻璃珠法拖尾严重。借助高通量测序方法检测,两种提取方法得到的样品基因组DNA在文库中的序列基本都能够被测出,在门和属水平上菌群结构相差不大。综上,改良CTAB法更适合大批量样品进行非培养方式的基因组DNA提取,该方法提取得到的基因组DNA浓度高且不影响后续实验。

To better study the microbial diversity in sufu,the genomic DNA was extracted from microorganisms in sufu without culture by modified cetyl trimethyl ammonium bromide(CTAB)method and glass bead method,and the extracted products were analyzed by high-throughput sequencing.The results showed that in terms of genomic DNA extraction concentration,both glass bead method and modified CTAB method could obtain a certain amount of genomic DNA and amplification bands,but the glass bead method had severe tailing.Through high-throughput sequencing method detection,the sequences of the sample genomic DNA obtained by two extraction methods in the library could be basically measured,and there was no large difference in the microbial community structure of sample at phylum and genus levels.Overall,the modified CTAB method was more suitable for genome DNA extraction of large quantities of samples without culture,and the concentration of genomic DNA extracted by this method was high and didn't affect subsequent experiments.