[1]何海燕,沙伟,张艳馥.大豆ISSR-PCR反应体系的优化[J].大豆科学,2010,29(03):510-513.[doi:10.11861/j.issn.1000-9841.2010.03.0510]
　HE Hai-yan,SHA Wei,ZHANG Yan-fu.Optimization for ISSR-PCR Reaction System of Soybean[J].Soybean Science,2010,29(03):510-513.[doi:10.11861/j.issn.1000-9841.2010.03.0510]
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大豆ISSR-PCR反应体系的优化

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第29卷 期数: 2010年03期 页码: 510-513 栏目: 出版日期: 2010-06-25
Title:
Optimization for ISSR-PCR Reaction System of Soybean
文章编号:
1000-9841（2010）03-0510-04
作者:
何海燕; 沙伟; 张艳馥
齐齐哈尔大学 生命科学与工程学院，黑龙江 齐齐哈尔 161006
Author(s):
HE Hai-yan; SHA Wei; ZHANG Yan-fu
College of Life Science and Engineering, Qiqihar University, Qiqihar 161006,Heilongjiang，China
关键词:
大豆; ISSR-PCR; 体系优化
Keywords:
Soybeen; ISSR-PCR; System optimization
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2010.03.0510
文献标志码:
A
摘要:
以黑龙江省大豆为材料，利用正交试验设计，对大豆ISSR-PCR反应体系中的5种主要因素(Mg2+、引物、dNTPs、模板DNA量、Taq酶量)在4个水平上进行体系优化。结果确定了大豆ISSR-PCR反应的最佳体系(25 μL)为：Mg2+浓度1.85 mmol·L-1、dNTPs浓度1.2mmol?L-1、引物浓度1.2μmol?L-1、模板DNA60 ng、Taq酶量0.7 U。利用该最佳体系，选取引物855对25份材料进行扩增，以验证该体系的稳定性。建立了适于大豆的ISSR-PCR反应体系，为利用ISSR标记技术开展黑龙江省大豆遗传多样性分析提供了依据。
Abstract:
Orthogonal design was adopted to optimize ISSR-PCR amplification system on soybean in five factors(Mg2+,dNTPs,primer,DNA template and Taq DNA polymerase)at four levels respectively, with 25 soybean germplasm from Heilongjiang Province as material. The most suitable ISSR-PCR system for soybean was established, namely 25 μL reaction system containing 1.85 mmol·L-1Mg2+,1.2mmol·L-1dNTPs,60 ng DNA template, 1.2μmol·L-1primer, and 0.7 U Taq DNA polymerase. Underthe optimized reaction conditions, 25 soybean were easily amplified with primer 855. The result provided a standardizing program for the analysis of interspecies genetic diversity of soybean of Heilongjiang Province.

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备注/Memo

基金项目：黑龙江省教育厅科研资助项目(10551331)。?

第一作者简介：何海燕 (1985-)，女，在读硕士，现从事植物遗传学研究。 E-mail：hehaiyan122@163.com。
通讯作者：沙伟，教授，博士生导师。E-mail：Shw1129@263.net。

更新日期/Last Update: 2014-09-14