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Evaluation of DNA Extraction and Purification Methods for Corn Straw Biogas Slurry
Qiao, J. T; Guo, R. B; Shi, X. S; Qiu, Y. L.
2013
发表期刊Advanced Materials Research
期号773页码:113-117
摘要Corn straw biogas slurry always contains humic substances, which poses particular
challenges in obtaining PCR-amplifiable DNA for analysis of microbial community. To establish an efficient and reliable DNA extraction method for straw biogas slurry, four approaches: i.e., direct SDS-based method, direct bead-based method, indirect SDS-based method, and indirect bead-based method were evaluated by comparing DNA yield, humic acid contamination, PCR amplifiability, and restriction fragment length polymorphisms (RFLP) of amplified 16S rRNA genes. Direct DNA extraction methods yielded 3-fold higher amounts of DNA than indirect procedures, but its DNA purity was lower. The A260/A230 ratio of DNA from indirect methods (0.8-0.85) were higher than that of DNA from direct methods (0.5-0.6), indicating DNA from direct methods contained high levels of humate contamination. PCR amplification was successful with crude DNA from indirect methods, but not with crude DNA from direct methods. PCR products could also be obtained with purified DNA from direct bead-based method, whereas not direct SDS-based method. Among the four methods, direct bead-based method, indirect SDS-based method and indirect bead-based method could obtain high-quality DNA extracts from corn straw biogas slurry. RFLP analysis further demonstrated the restriction patterns of amplified 16S rRNA genes from three methods were relatively identical microbial diversity.
关键词Corn Straw Biogas Slurry Molecular Ecology Dna Extraction Pcr Rflp
学科领域生物制氢与沼气
收录类别EI
语种英语
文献类型期刊论文
条目标识符http://ir.qibebt.ac.cn/handle/337004/1612
专题工业生物燃气团队
推荐引用方式
GB/T 7714
Qiao, J. T,Guo, R. B,Shi, X. S,et al. Evaluation of DNA Extraction and Purification Methods for Corn Straw Biogas Slurry[J]. Advanced Materials Research,2013(773):113-117.
APA Qiao, J. T,Guo, R. B,Shi, X. S,&Qiu, Y. L..(2013).Evaluation of DNA Extraction and Purification Methods for Corn Straw Biogas Slurry.Advanced Materials Research(773),113-117.
MLA Qiao, J. T,et al."Evaluation of DNA Extraction and Purification Methods for Corn Straw Biogas Slurry".Advanced Materials Research .773(2013):113-117.
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