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Rapid extraction and preservation of genomic DNA from human samples

机译:从人类样品中快速提取和保存基因组DNA

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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. [Figure not available: see fulltext.]
机译:简单快速提取人类基因组DNA仍然是基因组分析和疾病诊断的瓶颈。当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。我们报告了从颊拭子和唾液样品中提取人类基因组DNA的新方法。通过电场和毛细作用将DNA吸引到镀金的微芯片上,同时将捕获的DNA通过70°C的加热加热洗脱。设计了一种原型设备来处理四个微芯片,并开发了兼容的协议。使用微芯片通过不同长度的PCR扩增子,核和线粒体基因的不同长度的qPCR对使用微芯片提取的DNA进行表征。与市售试剂盒相比,只需较少的步骤即可获得相等的DNA提取产量。对捕获的DNA进行了室温保存1个月的证明,以一种环境友好的方案促进了基因组DNA的直接收集,传递和处理。 [图不可用:请参见全文。]

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