[1]黎丽倩,李妮亚,刘强,等. 假臭草ISSR-PCR反应体系的建立与优化[J].西北林学院学报,2013,28(02):83-89.[doi:10.3969/j.issn.1001-7461.2013.02.16]
　LI Li-qian,LI Ni-ya,LIU Qiang,et al. Establishment and Optimization of ISSR-PCR Reaction System for Eupatorium catarium[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2013,28(02):83-89.[doi:10.3969/j.issn.1001-7461.2013.02.16]
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 假臭草ISSR-PCR反应体系的建立与优化()
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《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]

卷:
第28卷
期数:
2013年02期
页码:
83-89
栏目:
出版日期:
2013-03-30

文章信息/Info

Title:
 Establishment and Optimization of ISSR-PCR Reaction System for Eupatorium catarium
文章编号:
1001-7461(2013)02-0083-07
作者:
 黎丽倩; 李妮亚; 刘强; 王婷; 杨丽; 石俊菲
 (热带动植物生态学省部共建教育部重点实验室 海南师范大学 生命科学学院，海南 海口 571158)
Author(s):
 LI Li-qian; LI Ni-ya; LIU Qiang; WANG Ting; YANG Li; SHI Jun-fei
 (Key Laboratory for Tropical Animal and Plant Ecology,Ministry of Education，College of Life Science,Hainan Normal University,Haikou,Hainan 571158,China)
关键词:
 ; 假臭草; ISSR; 反应体系; 优化
Keywords:
Eupatorium catarium;  ISSR;  reaction system;  optimization
分类号:
S722.33
DOI:
10.3969/j.issn.1001-7461.2013.02.16
文献标志码:
A
摘要:
 采用单因子试验、L16 (45)正交试验设计，以假臭草DNA为模板，研究了PCR反应体系中的5种主要成分，即Mg2+、dNTPs、Taq酶、引物及模板，对假臭草ISSR-PCR扩增结果的影响，并对引物退火温度进行了筛选和优化。建立了假臭草ISSR最佳反应体系：在20 μL的反应体系中，Mg2+为2.0 mmol/L，模板DNA用量为60 ng,TaqDNA聚合酶为1.5 U,dNTPs浓度为0.2 mmol/L，引物浓度为0.3 μmol/L。假臭草ISSR反应体系的建立为利用该技术分析假臭草遗传多样性及探索防控措施提供良好的基础。
Abstract:
 The effects of 5 factors ( Mg2+ concentration,dNTPs concentration,Taq DNA polymerase dosage,primer concentration and DNA templates concentration) on ISSR amplification were tested,and the annealing temperature was selected and optimized for establishment of a suitable ISSR-PCR reaction system of Eupatorium catarium through single factor test and orthogonal experiment design of L16 (45).The results showed that the optimal ISSR-PCR reaction system (20 μL) contained 2.0 mmol/L Mg2+，60 ng DNA template，1.5 U Taq DNA polymerase，200 μmol/L dNTPs and 0.3 μmol/L primer.The establishment of the ISSR-PCR reaction system could lay a favorable foundation for the analysis of the genetic diversity of E.catarium and for the exploration of prevention and control strategy for E.catarium.

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备注/Memo

备注/Memo:
 收稿日期：2012-07-07 修回日期：2012-10-12
基金项目：国家自然科学基金项目（31060095）。
作者简介：黎丽倩，女，硕士研究生，主要研究方向:恢复生态学。E-mail:395005396@qq.com
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