Rapid extraction and preservation of genomic DNA from human samples.
Rapid extraction and preservation of genomic DNA from human samples.
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从人类样本中快速提取和保存基因组 DNA。
DOI：
10.1007/s00216-012-6637-8
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发表时间：
2013
影响因子：
4.3
通讯作者：
Chung,J-H
中科院分区：
文献类型：
--
作者：
Kalyanasundaram,D;Kim,J-H;Yeo,W-H;Oh,K;Lee,K-H;Kim,M-H;Ryew,S-M;Ahn,S-G;Gao,D;Cangelosi,GA;Chung,J-H
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。目前使用微过滤器的方法需要繁琐的多个处理步骤，部分原因是必须控制盐条件，以吸引和洗脱多孔二氧化硅中的DNA。我们报道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镀金的微芯片上，而捕获的DNA通过70℃的加热洗脱。设计了一个处理四个微芯片的原型装置，并开发了兼容的协议。对不同样本量、不同长度的扩增产物、核基因和线粒体基因进行定量聚合酶链式反应(QPCR)分析。与商业试剂盒相比，提取DNA的步骤更少，提取得率相同。捕获的DNA在室温下保存了1个月，便于以环境友好的协议直接收集、传递和处理基因组DNA。图便携式人类基因组DNA提取微尖设备
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.FigurePortable microtip device for human genomic DNA extraction