| 栝楼的RAPD-PCR体系建立与优化 |
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| 引用本文： | 曲益涛,董玲,江芹,宁志怨,廖华俊. 栝楼的RAPD-PCR体系建立与优化[J]. 安徽农业科学, 2010, 38(13): 7150-7152. DOI: 10.3969/j.issn.0517-6611.2010.13.036 |
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| 作者姓名： | 曲益涛 董玲 江芹 宁志怨 廖华俊 |
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| 作者单位： | 安徽农业大学园艺学院,安徽合肥,230036;安徽省农业科学院,安徽合肥,230031 |
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| 摘 要： | [目的]建立栝楼的RAPD-PCR体系并对该体系进行优化。[方法]以栝楼叶片为材料,采用CTAB法提取栝楼叶片基因组DNA,利用正交设计对RAPD-PCR体系进行优化。[结果]各因素水平变化对PCR反应影响大小依次为：Mg2＋、TaqDNA聚合酶、引物和dNTPs。通过试验分析,优化的栝楼RAPD反应体系为：在25μl反应体系中,含10×buffer2.5μl,Mg2＋2.0mmol/L,Taq酶1U,引物0.8μmol/L,dNTPs0.1mmol/L。反应程序为94℃预变性2mim;94℃变性30s,37℃退火温度40s,72℃延伸1.5mim,36次循环;72℃延伸10mim,4℃保存。[结论]该研究建立的栝楼RAPD反应体系稳定可靠,为栝楼性别鉴定、遗传多样性分析、亲缘关系分析等方面的研究提供了有效的方法。
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| 关 键 词： | 栝楼 RAPD 分子标记 优化 |
Establishment and Optimization of RAPD-PCR System in Trichosanthes kirilowii Maxim |
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| QU Yi-tao et al. Establishment and Optimization of RAPD-PCR System in Trichosanthes kirilowii Maxim[J]. Journal of Anhui Agricultural Sciences, 2010, 38(13): 7150-7152. DOI: 10.3969/j.issn.0517-6611.2010.13.036 |
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| Authors: | QU Yi-tao et al |
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| Affiliation: | QU Yi-tao et al (Anhui Agricultural University,Hefei,Anhui 230036) |
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| Abstract: | [Objective] The aim was to establish and optimize the RAPD-PCR system of Trichosanthes kirilowii Maxim. [Method] With the leaves of T. kirilowii as materials, the genomic DNA of T. kirilowii leaves were extracted by CTAB method and the orthogonal design was used to optimize the RAPD-PCR system of T. kirilowii. [Result] The effects of level changes of each factor on PCR reaction in the following order was Mg2+, Taq DNA polymerase, primer and dNTPs. Through experimental analysis, the optimized RAPD-PCR system... |
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| Keywords: | RAPD |
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