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用于通过聚合酶链式反应(PCR)分析的植物组织制备的单步方案。

Single-step protocol for preparation of plant tissue for analysis by PCR.

作者信息

Thomson D, Henry R

机构信息

Queensland Agricultural Biotechnology Centre, St. Lucia, Australia.

出版信息

Biotechniques. 1995 Sep;19(3):394-7, 400.

PMID:7495552
Abstract

PCR has many applications in the isolation and analysis of plant DNA. The influence of salt and EDTA concentration, pH, incubation time and temperature on the preparation of plant material for PCR was evaluated. A general single-step method was developed in which a small amount of plant tissue was heated in a simple solution. The DNA in the supernatant was found to be suitable for most PCR applications including arbitrarily primed PCR (random-amplified polymorphic DNA) and PCR with specific primers for both single- and multiple-copy genes. The technique is much simpler than those generally used for plant DNA preparation and was successful with tissues from a wide range of species.

摘要

聚合酶链式反应(PCR)在植物DNA的分离与分析中有许多应用。评估了盐浓度、乙二胺四乙酸(EDTA)浓度、pH值、孵育时间和温度对用于PCR的植物材料制备的影响。开发了一种通用的单步方法,即把少量植物组织在一种简单溶液中加热。发现上清液中的DNA适用于大多数PCR应用,包括任意引物PCR(随机扩增多态性DNA)以及使用针对单拷贝和多拷贝基因的特异性引物进行的PCR。该技术比通常用于植物DNA制备的技术要简单得多,并且在多种植物的组织上都取得了成功。

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