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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
Source PublicationAdvanced Materials Research
Issue773Pages:113-117
AbstractCorn 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.
KeywordCorn Straw Biogas Slurry Molecular Ecology Dna Extraction Pcr Rflp
Subject Area生物制氢与沼气
Indexed ByEI
Language英语
Document Type期刊论文
Identifierhttp://ir.qibebt.ac.cn/handle/337004/1612
Collection工业生物燃气研究组
Recommended Citation
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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