大连理工大学学报
大連理工大學學報
대련리공대학학보
JOURNAL OF DALIAN UNIVERSITY OF TECHNOLOGY
2014年
3期
272-277
,共6页
王晓辉%黄李淑馨%白凤武%杜昱光
王曉輝%黃李淑馨%白鳳武%杜昱光
왕효휘%황리숙형%백봉무%두욱광
宏基因组DNA%提取%纯化%海洋底泥
宏基因組DNA%提取%純化%海洋底泥
굉기인조DNA%제취%순화%해양저니
metagenomic DNA%extraction%purification%marine sediment samples
为了探索有效的海洋底泥宏基因组DNA提取与纯化方法,分别采用CTAB结合SDS法、CTAB结合玻璃珠法、SDS结合蛋白酶 K 法和 SoilMasterTM试剂盒法提取海洋底泥宏基因组DNA,用柱式腐殖酸清除试剂盒法与电洗脱法纯化宏基因组 DNA,并对宏基因组 DNA的产量与纯度进行评价。结果表明:SDS 结合蛋白酶 K 法提取率最高,DNA 片段完整;SoilMasterTM试剂盒法与 CTAB结合 SDS法次之;CTAB结合玻璃珠法有严重降解;电洗脱法可以得到高纯度、大于42 kb宏基因组DNA片段。因此,SDS结合蛋白酶 K 法和电洗脱法提取纯化海洋近海底泥宏基因组DNA优于其他方法。
為瞭探索有效的海洋底泥宏基因組DNA提取與純化方法,分彆採用CTAB結閤SDS法、CTAB結閤玻璃珠法、SDS結閤蛋白酶 K 法和 SoilMasterTM試劑盒法提取海洋底泥宏基因組DNA,用柱式腐殖痠清除試劑盒法與電洗脫法純化宏基因組 DNA,併對宏基因組 DNA的產量與純度進行評價。結果錶明:SDS 結閤蛋白酶 K 法提取率最高,DNA 片段完整;SoilMasterTM試劑盒法與 CTAB結閤 SDS法次之;CTAB結閤玻璃珠法有嚴重降解;電洗脫法可以得到高純度、大于42 kb宏基因組DNA片段。因此,SDS結閤蛋白酶 K 法和電洗脫法提取純化海洋近海底泥宏基因組DNA優于其他方法。
위료탐색유효적해양저니굉기인조DNA제취여순화방법,분별채용CTAB결합SDS법、CTAB결합파리주법、SDS결합단백매 K 법화 SoilMasterTM시제합법제취해양저니굉기인조DNA,용주식부식산청제시제합법여전세탈법순화굉기인조 DNA,병대굉기인조 DNA적산량여순도진행평개。결과표명:SDS 결합단백매 K 법제취솔최고,DNA 편단완정;SoilMasterTM시제합법여 CTAB결합 SDS법차지;CTAB결합파리주법유엄중강해;전세탈법가이득도고순도、대우42 kb굉기인조DNA편단。인차,SDS결합단백매 K 법화전세탈법제취순화해양근해저니굉기인조DNA우우기타방법。
In order to seek an effective extraction and purification method of metagenomic DNA from marine sediment samples,the metagenomic DNA was extracted by CTAB plus SDS method,CTAB plus glass beads method,SDS plus proteinase K method and SoilMasterTM DNA Extraction Kit method,respectively.The metagenomic DNA was purified by column humic clear kit method and electroelution.The quality and quantity of the extracted metagenomic DNA are compared.The highest extracted rate and integrity are obtained by SDS plus proteinase K method, next are SoilMasterTM DNA Extraction Kit method and CTAB plus SDS method;CTAB plus glass beads method has severe degradation;and electroelution produces the purest metagenomic DNA and large fragments with more than 42 kb.The research results show that SDS plus proteinase K method and electroelution are the optimum extraction and purification methods of metagenomic DNA from marine sediment samples compared with other methods.