适用于川芎的ISSR-PCR反应体系的建立及优化 |
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引用本文: | 王岚,唐琳.适用于川芎的ISSR-PCR反应体系的建立及优化[J].安徽农业科学,2011,39(23):13949-13951. |
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作者姓名: | 王岚 唐琳 |
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作者单位: | 四川大学生命科学学院,四川成都610064;中州大学化工食品学院,河南郑州450044;四川大学生命科学学院,四川成都,610064 |
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基金项目: | “十五”国家科技攻关项目(2004BA721A31) |
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摘 要: | 目的]建立和优化适合川芎的ISSR-PCR反应体系。方法]以川芎叶片为材料,利用改良CTAB法提取了叶片基因组DNA,利用单因素试验分析了DNA模板、Mg2+、dNTP、引物和TaqDNA聚合酶的浓度对ISSR-PCR反应的影响,对适合川芎的ISSR-PCR反应条件进行了优化。结果]适合川芎的ISSR-PCR反应体系为25.0μl,其中含2.5μl 10×TaqDNA聚合酶缓冲液、2.1 mmol/LMgC l2、300μmol/L dNTP、0.4μmol/L引物、1.0 UTaqDNA聚合酶和20~40 ng模板DNA。结论]为进一步对全国17个不同分布区的川芎资源进行遗传多样性研究奠定了基础。
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关 键 词: | 川芎 ISSR-PCR 体系优化 |
Establishment and Optimization of ISSR-PCR Amplification System for Ligusticum chuanxiong Hort |
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Institution: | WANG Lan et al(College of Life Sciences,Sichuan University,Chengdu,Sichuan 610064) |
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Abstract: | Objective] This study was to establish and optimize the ISSR-PCR reaction system for Ligusticum chuanxiong Hort.Method] Using genomic DNA of Chuanxiong leaf extracted via an improved CTAB method,single factor analysis was performed to investigate the impacts of DNA template concentration,Mg2+ concentration,dNTPs concentration,primer concentration,Taq DNA polymerase concentration on ISSR-PCR amplification and to optimize this system for Ligusticum chuanxiong Hort.Result] The ISSR-PCR amplification(25 μl) suitable for Ligusticum chuanxiong Hort.was determined to be composed of 2.5 μl of 10 × reaction buffer,2.1 mmol/L MgCl2,300 μmol/L dNTPs,0.4 μmol/L primer,1.0 U Taq DNA polymerase and 20-40 ng genomic DNA.Conclusion] Our study laid basis for analyzing the genetic diversity of Ligusticum chuanxiong Hort.resources distributed in 17 different areas of China. |
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Keywords: | Ligusticum chuanxiong Hort ISSR-PCR Parameter optimization |
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