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从人体样本中快速提取和保存基因组 DNA。

Rapid extraction and preservation of genomic DNA from human samples.

机构信息

Department of Mechanical Engineering, University of Washington, Seattle, WA 98195, USA.

出版信息

Anal Bioanal Chem. 2013 Feb;405(6):1977-83. doi: 10.1007/s00216-012-6637-8. Epub 2013 Jan 11.

Abstract

Simple and rapid extraction of human genomic DNA remains a bottleneck for genome analysis and disease diagnosis. Current methods using microfilters require cumbersome, multiple handling steps in part because salt conditions must be controlled for attraction and elution of DNA in porous silica. We report a novel extraction method of human genomic DNA from buccal swab and saliva samples. DNA is attracted onto a gold-coated microchip by an electric field and capillary action while the captured DNA is eluted by thermal heating at 70 °C. A prototype device was designed to handle four microchips, and a compatible protocol was developed. The extracted DNA using microchips was characterized by qPCR for different sample volumes, using different lengths of PCR amplicon, and nuclear and mitochondrial genes. In comparison with a commercial kit, an equivalent yield of DNA extraction was achieved with fewer steps. Room-temperature preservation for 1 month was demonstrated for captured DNA, facilitating straightforward collection, delivery, and handling of genomic DNA in an environment-friendly protocol.

摘要

从口腔拭子和唾液样本中简单快速地提取人类基因组 DNA 仍然是基因组分析和疾病诊断的瓶颈。目前使用微滤器的方法需要繁琐的、多次处理的步骤,部分原因是盐条件必须控制以吸引和洗脱多孔二氧化硅中的 DNA。我们报告了一种从口腔拭子和唾液样本中提取人类基因组 DNA 的新方法。DNA 通过电场和毛细作用被吸引到金涂覆的微芯片上,而捕获的 DNA 则通过在 70°C 下热加热洗脱。设计了一种用于处理四个微芯片的原型设备,并开发了兼容的方案。使用微芯片提取的 DNA 针对不同的样本体积、不同长度的 PCR 扩增子以及核基因和线粒体基因进行了 qPCR 表征。与商业试剂盒相比,该方法提取 DNA 的步骤更少,产量相当。还证明了捕获的 DNA 在室温下保存 1 个月,方便在环保方案中直接收集、运输和处理基因组 DNA。

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