中国海洋大学学报（自然科学版）2017，Vol.47Issue(6)：17-24,8.DOI:10.16441/j.cnki.hdxb.20160246
海洋沉积物中微生物基因组DNA提取方法的比较研究
Comparison Study on Extraction Methods of Microbial DNA in Marine Sediments
摘要
Abstract
The composition of sediments is complex, and the impurities, such as humic acid, are difficult to remove when DNA extraction.Because of the inhibition on PCR amplification, high quality DNA is the basis and prerequisite for the molecular ecology of marine microorganisms.In this study, six different methods were used to extract microbial DNA from marine sediments.Comparisons of the purity and yield of microbial DNA, 16S rRNA gene copy numbers and the characteristics of microbial community were used to analyze the quality and quantity of the genomic DNA.The results showed that DNA purity obtained by method 1, method 2 and method 3 were high despite low in yield, which could be directly used in the following microbial ecology analysis.Compared to method 2, the bacterial 16S rRNA gene copy numbers and template amount for bacterial 16S rRNA gene in per nanogramme DNA obtained by method 3 was low.DNA obtained by method 4 was high in yield but low in purity.After purification (method 5 and method 6), DNA samples were high in purity but there were massive losses.Moreover, method 2 could fully break up cells, reflecting the characteristics of microbial community better.Synthesizing the above results, the method 2 (PowerSoil DNA Isolation Kit) was the best one to marine sediment microbial DNA extraction.This study provides an effective technique for the molecular ecology of marine microorganisms.关键词
沉积物/微生物/DNA提取Key words
sediments/microbe/DNA extraction分类
生物科学引用本文复制引用
贺惠,张玉,甄毓,米铁柱,陈阳阳,于志刚..海洋沉积物中微生物基因组DNA提取方法的比较研究[J].中国海洋大学学报（自然科学版）,2017,47(6):17-24,8.基金项目
国家自然科学基金项目(41521064 （）
41376093) （）
中国科学院海洋生态与环境科学重点实验室、青岛海洋科学与技术国家实验室海洋生态与环境科学开放课题项目(KLMEES201601)资助 Supported by the National Natural Science Foundation of China(41521064 （KLMEES201601）
41376093) （）
the Open Fund of Key Laboratory of Marine Ecology and Environmental Sciencea,Institute of Oceanology,Chinese Academy of Sciences,and Laboratory of Marine Ecology and Environmental Science,Qingdao National Laboratory for Marine Science and Technology(KLMEES201601) （KLMEES201601）
