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从叶苔科植物附叶平叶苔中改进基因组DNA提取和PCR扩增的方案

Protocol for improved extraction and PCR amplification of genomic DNA from liverwort, Plagiochasma appendiculatum.

作者信息

Soni Arvind, Kumar Anil

机构信息

Plant Genomic Lab, Molecular Biology and Genetic Engineering Division, National Botanical Research Institute, Luckonw 266 001, India.

出版信息

Indian J Exp Biol. 2009 Nov;47(11):921-4.

Abstract

A simplest method was followed for isolation of high quality genomic DNA form thallus of Plagiochasma appendiculatum that contained large quantities of polyphenols, terpenoids, tannins, contamination of high amount of RNA and polysaccharides. The method involved a modification of CTAB procedure using PVP (1%) and LiCl (4M) solution to remove polyphenols and RNA and some other binding proteins. The present protocol was found suitable for restriction enzyme digestion and random amplified polymorphic DNA (RAPD) analysis.

摘要

采用了一种最简单的方法从具尾裂叶苔的叶状体中分离高质量的基因组DNA,该叶状体含有大量的多酚、萜类、单宁、大量RNA和多糖污染。该方法涉及对CTAB程序进行修改,使用聚乙烯吡咯烷酮(1%)和氯化锂(4M)溶液去除多酚、RNA和其他一些结合蛋白。发现本方案适用于限制性内切酶消化和随机扩增多态性DNA(RAPD)分析。

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