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海洋底泥环境DNA提取与纯化方法比较 |
Comparative Study of Methods for Extraction and Purificationof Environmental DNA From Marine Sediment Samples |
投稿时间:2013-07-17 修订日期:2013-07-17 |
DOI: |
中文关键词: 环境DNA,提取,纯化,海洋底泥 |
英文关键词: Environmental DNA, extraction, Purification, marine sediments |
基金项目:国家高技术研究发展计划(863计划)课题“化学品生物转化过程与装备(2012AA021205)”;中国科学院知识创新工程重要方向课题“果糖基平台化合物及下游产品关键制备技术(KSCX2-EW-G-5)”;国家科技支撑计划课题“大型海藻新型产品加工技术(2013BAB01B01)” |
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中文摘要: |
[目的]寻找有效的海洋底泥环境DNA提取与纯化方法。[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、试剂盒法提取海洋底泥宏基因组DNA;分别采用试剂盒与电洗脱法纯化环境DNA,比较评价其质量与纯度。[结果] 蛋白酶K+SDS法提取率最高、片段完整性最好;试剂盒法与CTAB结合SDS法次之;CTAB结合玻璃珠法降解严重;电洗脱法纯化出高纯度、高浓度、大于42kb片段。[结论]从经济和DNA质量的角度考虑,蛋白酶K+SDS法和电洗脱法作为提取纯化海洋底泥环境DNA的最佳方法。 |
英文摘要: |
[Objective] The aim was to seek an effective extraction ang purification method of metagenomic DNA from marine sediment. [Method] The metagenmic DNA was extracted by CTAB method、SDS-Proteinase K method、kit method, respectively, which was purified by kit and electroelution. Quality and quantity of the extracted DNA were compared. [Result] the extracted rate and integrity by SDS combined with protease K method were the highest, next was kit method and CTAB combined with SDS method; Electroelute purified environmental DNA produced the purest, fragments are larger in size (>42kb). [Conclusion]Considering economy and DNA quality, SDS combined with protease K method and Electroelution regarded as the optimum method of metagenomic DNA from marine sediment. |
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