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通过缠绕法分离涡虫高分子量DNA

Planarian High Molecular Weight DNA Isolation by Spooling.

作者信息

Zhang Shasha, Alvarado Alejandro Sánchez

机构信息

Howard Hughes Medical Institute, Stowers Institute for Medical Research, Kansas City, MO, USA.

出版信息

Methods Mol Biol. 2018;1774:277-284. doi: 10.1007/978-1-4939-7802-1_8.

Abstract

High-quality large molecular weight genomic DNA is important for genomic studies. Most commercial available genomic DNA purification kits have failed to generate high molecular weight DNA of sufficient quality from planarians. Here, we describe a simple and efficient genomic DNA isolation method, which has worked for several different planarian species, including Schmidtea mediterranea. This phenol-chloroform based method can be used to obtain genomic DNA of up to 150 kb and can be used for bacterial artificial chromosome (BAC) library construction, next-generation sequencing and PCR cloning.

摘要

高质量的大分子基因组DNA对于基因组研究很重要。大多数市售的基因组DNA纯化试剂盒都未能从涡虫中获得足够质量的高分子量DNA。在此,我们描述了一种简单有效的基因组DNA分离方法,该方法已适用于几种不同的涡虫物种,包括地中海涡虫。这种基于酚-氯仿的方法可用于获得长达150 kb的基因组DNA,可用于细菌人工染色体(BAC)文库构建、下一代测序和PCR克隆。

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