Establishment and Optimization of ISSR Reaction System for Carpetgrass
Establishment and Optimization of ISSR Reaction System for Carpetgrass
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DOI：
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发表时间：
2004
期刊：
影响因子：
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通讯作者：
J.-B. Xi;Yuzhong Zheng;Zhongyi Yang
中科院分区：
文献类型：
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作者：
J.-B. Xi;Yuzhong Zheng;Zhongyi Yang
对地毯草遗传多样性研究中影响ISSR分析的因素，如基因组DNA提取、模板DNA浓度、Taq DNA聚合酶、Mg2+浓度、dNTP和退火温度等进行了研究，优化了ISSR-PCR的反应条件。结果表明，适合地毯草ISSR-PCR反应的条件为：1 × Taq缓冲液（10mmol/LTris-HCl，pH90，50mmol/LKCl，0.1%TritonX-100），0.75UTaqDNA（申能公司生产），0.375 μ mol/L引物，180 μ mol/LdNTP，15~2.0mmol/LMgCl2，10ng模板DNA。
Factors which affect the ISSR analysis in the study of the genetic diversity of carpetgrass, such as genomic DNA extraction, concentration of template DNA, Taq DNA polymerase, Mg~(2+) concentration, dNTP and annealing temperature, were studied for optimizing conditions of the ISSR-PCR. The results showed that the conditions being suitable for ISSR-PCR of carpetgrass were as follows: 1×Taq buffer (10 mmol/L Tris-HCl, pH 9\^0, 50 mmol/L KCl, 0.1% Triton X-100),0.75 U Taq DNA (produced by Shenneng company), 0.375 μmol/L primer, 180 μmol/L dNTP, 1\^5～2.0 mmol/L MgCl\-2, and 10 ng template DNA.