| 亚麻RAPD-PCR体系的优化及引物的筛选 |
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| 引用本文： | 王福亮,黄文功.亚麻RAPD-PCR体系的优化及引物的筛选[J].安徽农业科学,2009,37(15):6911-6913. |
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| 作者姓名： | 王福亮 黄文功 |
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| 作者单位： | 黑龙江省农业科学院原子能研究所,黑龙江,哈尔滨,150086;黑龙江省农业科学院经济作物研究所,黑龙江,哈尔滨,150086 |
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| 摘 要： | 目的]优化亚麻RAPD反应条件，筛选亚麻RAPD引物。方法]DNA条带扩增采用PCR技术，条带的分离采用琼脂糖凝胶电泳法，成像利用紫外成像系统。结果]试验优化亚麻RAPD反应条件：25出反应体系中，含10×Buffer，Mg^2＋ （1mmol／L），Taq酶1U，Genome DNA 50ng．dNTP 0．25mmol／L．RAPD primer1．5μmol／L。PER反应程序为：94℃预变性4min；97℃变性40s，37℃退火1min，72℃延伸90s，40个循环；72℃延伸10min。试验利用3个有代表性的亚麻品种从70条引物中筛选出12条多态性好，重复性高的引物。结论]该试验筛选出12条引物，并优化了亚麻RAPD反应条件，为亚麻的分子鉴定奠定了基础。
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| 关 键 词： | 亚麻 基因组DNA RAPD反应 筛选 |
Optimizing the RAPD-PCR System and Screening RAPD Primers for Flax |
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| Institution: | WANG Fu-liang et al (Institute of Atomic Energy, Heilongjiang Academy of Agricultural Sciences, Harbin, Heilo ngjiang 150086) |
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| Abstract: | Objective]This experiment aimed to optimize the RAPD reaction conditions of flax and screen RAPD primers of flax.Methods] DNA strip was amplified by PCR technology.And the strip was separated by agrose gel electrophoresis.The images were formed by using UV imaging system.Result] The RAPD reaction conditions of flax were optimized as following: in 25 μl reaction system,10×Buffer,Mg2+(1 mmol/L),Taq enzyme 1 U,Genome DNA 50 ng,dNTP 0.25 mmol/L and RAPD primer 1.5 μmol/L.The process of PCR were as follows: predegenerating at 94 ℃ for 4 min,40 cycles(degenerating at 94 ℃ for 40 s,annealing at 37 ℃ for 1 min,extending at 72 ℃for 90 s),extending at 72 ℃ for 10 min.Three typical flax varieties were used to screen out 12 primers with high level polymorphism and repeatability from 70 primers.Conclusion] 12 primers were screened out and RAPD reaction conditions of flax were optimized.This research laid the foundation for the molecular identification of flax. |
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| Keywords: | Flax Genome DNA RAPD reaction Screening |
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