[1]宋显军,谢甫绨,张伟,等.大豆SSR检测体系的优化研究[J].大豆科学,2007,26(01):36-40.[doi:10.3969/j.issn.1000-9841.2007.01.009]
　SONG Xian-jun,XIE Fu-ti,ZHANG Wei,et al.OPTIMIZATION OF SSR DETECTION SYSTEM IN SOYBEAN[J].Soybean Science,2007,26(01):36-40.[doi:10.3969/j.issn.1000-9841.2007.01.009]
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大豆SSR检测体系的优化研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第26卷 期数: 2007年01期 页码: 36-40 栏目: 出版日期: 2007-02-25
Title:
OPTIMIZATION OF SSR DETECTION SYSTEM IN SOYBEAN
文章编号:
1000-9841(2007)01-0036-05
作者:
宋显军; 谢甫绨; 张伟; 曹萍; 郭玉华
沈阳农业大学农学院，沈阳 110161
Author(s):
SONG Xian-jun; XIE Fu-ti; ZHANG Wei; CAO Ping; GUO Yu-hua
College of Agronomy， Shenyang Agricultural University， Shenyang 110161
关键词:
大豆; SSR; 检测体系;  优化
Keywords:
Soybean; SSR; Detection system; Optimization
分类号:
S565.1
DOI:
10.3969/j.issn.1000-9841.2007.01.009
文献标志码:
A
摘要:
以大豆为试材，研究了大豆SSR检测体系中PCR扩增的各主要成分对检测结果的影响，确立了适宜的退火温度和反应体积，改进了PCR扩增程序和银染方法，进一步优化了适用于大豆的SSR检测体系。优化后的SSR反应体系为： 10×Buffer 1.00μL、2.00 ng/μL模板DNA、0.15 μmol/L SSR引物、150.00μmol/L dNTPs、0.50 U Taq酶、2.00mmol/L Mg2+，加ddH2O至终体积10.00μL。优化的PCR扩增程序为：94℃预变性3 min；94℃变性25s，47℃退火25s，72℃延伸30s，共35个循环；72℃延伸5min，4℃保存。优化后的检测体系更为经济有效。
Abstract:
Effects of major components of PCR amplification on soybean SSR detection results were studied. A SSR detection system suitable for soybean was established with improved PCR reaction system， PCR program and silver staining method. The optimized PCR amplification system was 10×Buffer 1.00μL， 2.00 ng/μL template DNA， 0.15 μmol/L SSR primer， 150.00μmol/L dNTPs， 0.50 U Taq enzyme， 2.00 mmoL/L Mg2+， adding ddH2O to terminal volume 10.00 μL. The optimized PCR cycles started at 94℃(3min)，followed by 35 cycles of 25s at 94℃，25s at 47℃ and 30s at 72℃， and then extended 5min at 72℃.The optimized soybean SSR detection system was proved more economic and useful.

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备注/Memo

基金项目：本研究得到辽宁省教育厅科技项目（05L3781）和辽宁省科学技术计划项目（2006201008）的资助

作者简介：宋显军（1980-），男，在读硕士，从事大豆数量遗传研究。E-mail:mrsong1192003@yahoo.com.cn

更新日期/Last Update: 2014-10-21