Quantitative and qualitative validations of a sonication-based DNA extraction approach for PCR-based molecular biological analyses
Quantitative and qualitative validations of a sonication-based DNA extraction approach for PCR-based molecular biological analyses
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DOI：
10.1016/j.ab.2016.01.003
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发表时间：
2016-05-15
影响因子：
2.9
通讯作者：
Yan, Han
中科院分区：
文献类型：
--
作者：
Dai, Xiaohu;Chen, Sisi;Yan, Han
本研究的目的是全面验证基于超声波的DNA提取方法，以期取代所谓的标准DNA提取方法-商业试剂盒方法。消化后的污泥样品中含有较高数量的腐植酸和蛋白质等聚合酶抑制物质的微生物细胞被用作实验替代品。获得了污泥样品的固液分离、DNA模板和抑制剂的稀释、基于PCR分析的最小模板以及对焦磷酸测序技术偏差分析的深入了解，证实了基于超声波的DNA提取方法的有效性。(C)2016 Elsevier Inc.保留所有权利。
The aim of this study was to comprehensively validate the sonication-based DNA extraction method, in hope of the replacement of the so-called 'standard DNA extraction method' - the commercial kit method. Microbial cells in the digested sludge sample, containing relatively high amount of PCR-inhibitory substances, such as humic acid and protein, were applied as the experimental alternatives. The procedure involving solid/liquid separation of sludge sample and dilution of both DNA templates and inhibitors, the minimum templates for PCR-based analyses, and the in-depth understanding from the bias analysis by pyrosequencing technology were obtained and confirmed the availability of the sonication-based DNA extraction method. (C) 2016 Elsevier Inc. All rights reserved.