蝗虫肠道微生物总DNA提取方法的比较
Comparative studies on the methods of bacterial genomic DNA extraction from grasshopper guts.
龙文敏，严二平，宣 涛，魏 华，庞小燕，赵立平，马恩波
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作者单位：山西大学应用生物学研究所
中文关键词：蝗虫，肠道微生物，DNA提取，PCR-DGGE
英文关键词：grasshopper，hindgut microflora，DNA extraction，PCR-DGGE
中文摘要：采用Bead beating法和QIAamp DNA stool mini kit法提取蝗虫肠道微生物总DNA，并对2种方法提取DNA的得率、完整性以及16S rRNA基因扩增产物的变性梯度凝胶电泳(denaturing gradient gel electrophoresis, DGGE)图谱等进行综合比较。结果表明，Bead beating法提取DNA的得率显著高于QIAamp DNA stool mini kit法(P=0.042)，而QIAamp DNA stool mini kit法提取DNA片段更完整。PCR-DGGE检测微生物多样性结果显示，QIAamp DNA stool mini kit法提取DNA所代表的微生物群落多样性略高于Bead beating法，但Mann-Whitley统计学检验表明用2种方法检测蝗虫肠道微生物多样性无显著差异(P=0.17)。因此在蝗虫肠道微生物群落多样性的检测中QIAamp DNA stool mini kit法具一定的优势，而Bead beating法同样适用。
英文摘要：The effectiveness of two DNA extraction methods, bead beating and QIAamp DNA stool mini kit for isolation of the total microbial DNA from grasshopper guts, were compared. The two methods were evaluated comprehensively for their yield, integrality and readiness for subsequent analysis by polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE) of the bacterial communities. The results showed that the hindgut DNA yield by bead beating method was significantly higher (
P<0.05) than that of QIAamp DNA stool mini kit method, while the QIAamp DNA stool mini kit method produced DNA of larger size. In PCR-DGGE analysis, Mann-Whitley test (
P=0.17) showed that DNA extracted with QIAamp DNA stool mini kit revealed a more diverse microbial community, but there was no significant difference between the two methods. Consequently, the two methods are both suitable for analyzing the microbial community in grasshopper guts.