【摘要】 提取河流沉积物中DNA是研究河流沉积物微生物多样性的难点之一,实验开发了一种酚与SDS的DNA提取方法,与OMEGA和BioDee 2种DNA提取试剂盒方法进行了比较,并通过DNA纯度、PCR扩增结合16S rDNA克隆文库检验对几种方法进行了评价。结果表明,OMEGA DNA提取试剂盒法方法简单,提取的DNA纯度较高,酚+SDS法用时较长,DNA提取量大,纯度较低。BioDee试剂盒提取量最小,虽然对PCR结果没有影响,但不能满足沉积物微生物多样性分析的要求。OMEGA DNA提取试剂盒法和酚+SDS法可分别用于PCR扩增、克隆文库、DGGE等的后续实验。更多还原

【Abstract】 DNA extraction of river sediment sample is one of the difficulties in the research of the river sediment microbial diversity.An extraction method of DNA,i.e.phenol with SDS measure was developed in this study,and was compared with the other two DNA extraction kits of OMEGA and BioDee.The above mentioned methods were evaluated by the results of DNA purity,PCR amplification and 16S rDNA clone library.The results showed that the OMEGA DNA extraction kit method was a simple way with high DNA purity.The phenol + SDS method obtained plenty of DNA,however a longer time was needed and with lower purity DNA.The extraction rate of BioDee extraction kit method was minimum,but no effect on the PCR results.However,the PCR production was not suitable to the next steps of the analysis of sediment microbial diversity.The above results showed that the DNA,which was obtained using OMEGA DNA extraction kit method and the phenol + SDS method,could be used for the next steps of PCR,16S rDNA clone library,DGGE and other molecular tests.更多还原