落叶松cDNA扩增因素优化组合的研究  

Studies on the excellent combination of PCR amplification factors of Larix Principis-ruppreechxii cDNA

作　　者：周怀军[1] 张洪武[1] 安连荣[1] 刘敏丽[1] 杨丽[1] 张晓曼[1] 左永忠[1] 

机构地区：[1]河北农业大学林业资源与工程学院,河北保定071000

出　　处：《生物技术》2002年第6期3-5,共3页Biotechnology

基　　金：河北省教委科研项目"落叶松抗冻蛋白及抗冻基因克隆的研究"( 2 0 0 1 1 3)

摘　　要：为了探讨落叶松cDNA扩增因素优化组合 ,以落叶松总RNA为模板逆转录为cDNA ,然后进行PCR扩增 ,采用四因素三水平正交试验设计 ,分析比较了几种因素对PCR扩增的影响。结果显示 ,主要影响因素是Taq酶和Mg2 +,随机引物和dNTP的影响较小 ,不同水平的影响作用是 :Taq酶以高水平 ,即用量为 2U ,Mg2 +以中低水平 ,即用量为 1 5- 2 0mmol L ,可以得到多而清晰的条带 ;反之 ,则扩增效果差。In the research,cDNA was synthesized from DNA free total RNA by performing reverse transcription.Then several factors about DD-PCR were analysed by orthogenal design test L 9(34)and to expected combination of PCR amplification factors of larix principis-ruppereechxii.The result showed that the inflaming order of factors is:Taq DNA Polymerase>Mg 2+>dNTP=decamer primer.The inflaming result of different levels is:when Taq DNA Polymerase is used with high level(2U),and Mg 2+ is used in middle or low level (1.5-2.0mmol/L),numerous and distinct PCR bands could be detected.On the contrary,the PCR result is not good.

关 键 词：落叶松 cDNA扩增因素 优化组合 

分 类 号：S718.46[农业科学—林学] S791