从褐藻中提取高质量的基因组DNA。

Extraction of high-quality genomic DNA from Ectocarpus.

作者信息

Coelho Susana M, Scornet Delphine, Rousvoal Sylvie, Peters Nick, Dartevelle Laurence, Peters Akira F, Cock J Mark

机构信息

UPMC Université Paris 06, The Marine Plants and Biomolecules Laboratory, UMR 7139, Station Biologique de Roscoff, 29682 Roscoff Cedex, France.

出版信息

Cold Spring Harb Protoc. 2012 Mar 1;2012(3):365-8. doi: 10.1101/pdb.prot067967.

DOI:10.1101/pdb.prot067967

PMID:22383638

Abstract

For some applications, such as genome sequencing and high-throughput genotyping with multiple markers, it is necessary to use high-quality genomic DNA. This article describes how to obtain several micrograms of high-quality, cesium chloride-purified DNA from 1 g of Ectocarpus filaments. We also recommend using DNA of this quality for quantitative RT-PCR control reactions. However, simpler, more rapid, kit-based methods are preferable for experiments that involve the treatment of large numbers of individuals, such as genotyping large populations with a small number of markers or PCR screening of large populations.

摘要

对于某些应用，如基因组测序和使用多个标记的高通量基因分型，有必要使用高质量的基因组DNA。本文介绍了如何从1克海带丝中获得几微克高质量的、经氯化铯纯化的DNA。我们还建议将这种质量的DNA用于定量RT-PCR对照反应。然而，对于涉及大量个体处理的实验，如用少量标记对大量群体进行基因分型或对大量群体进行PCR筛选，更简单、更快速的基于试剂盒的方法更为可取。