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Enrichment of DNA Sequencing Libraries from Samples Containing Small Amounts of Target DNA

机译:从含有少量靶DNA的样品中富集DNA测序文库

摘要

Provided herein is a method for capturing DNA molecules in solution. The method may comprise: extracting DNA from a sample that comprises endogenous DNA and environmental DNA to produce extracted DNA; ligating universal adaptors to the extracted DNA; hybridizing the extracted DNA, in solution, with affinity-tagged RNA probes generated by: in vitro transcribing a library of fragmented reference genomic DNA that has been ligated to an RNA promoter adaptor, in the presence of an affinity-tagged ribonucleotide; binding the product with a capture agent that is tethered to a substrate in the presence of RNA oligonucleotides that are complementary to the adaptors, thereby capturing the hybridized DNA molecules on the substrate; washing the substrate to remove any unbound DNA molecules; and releasing the captured DNA molecules. A kit for performing the method is also provided.
机译:本文提供了一种用于捕获溶液中DNA分子的方法。该方法可以包括:从包含内源DNA和环境DNA的样品中提取DNA以产生提取的DNA;将通用衔接子连接至提取的DNA;在溶液中,将提取的DNA与亲和标记的RNA探针杂交,该探针通过以下方式产生:在亲和标记的核糖核苷酸存在下,体外转录已连接至RNA启动子衔接子的片段化参考基因组DNA文库;在与衔接子互补的RNA寡核苷酸的存在下,将产物与捕获剂结合,该捕获剂被束缚在基质上,从而将杂交的DNA分子捕获在基质上;洗涤底物以去除任何未结合的DNA分子;并释放捕获的DNA分子。还提供了用于执行该方法的套件。

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