| 干用辣椒RAPD—PCR反应体系正交优化的研究 |
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| 引用本文： | 王伟,郑根昌,闫光照,陈炳艳,包凤利.干用辣椒RAPD—PCR反应体系正交优化的研究[J].内蒙古民族大学学报(自然科学版),2009,24(1):55-57. |
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| 作者姓名： | 王伟 郑根昌 闫光照 陈炳艳 包凤利 |
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| 作者单位： | 1. 内蒙古民族大学农学院,内蒙古,通辽,028043
2. 北京德农种业,北京,100101 |
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| 摘 要： | 以干用辣椒幼苗为材料，研究了干用辣椒RAPD分析过程中的Taq酶、Mg^2＋,dNTPs、引物、模板DNA浓度、退火温度等影响因素，建立适合干用辣椒RAPD反应的PCR体系，即25μL反应体系中含有解体Taq酶1．5U、Mg^2＋2．5mmol／L，dNTPs0．6mmol／L、引物0．8μmol／L、模板DNA60ng．扩增程序为：94℃预变性4min；94℃变性1min，37℃退火1min，72％延伸1．5min，40个循环；最后72℃副延伸5min．
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| 关 键 词： | 干用辣椒 RAPD—PCR 反应体系 正交优化 |
Orthogonal Optimization For RAPD-PCR Reaction System Of Dry Peppers |
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| WANG Wei,ZHENG Gen-chang,YAN Guang-zhao,CHEN Bing-yan,BAO Feng-li.Orthogonal Optimization For RAPD-PCR Reaction System Of Dry Peppers[J].Journal of Inner Mongolia University for the Nationalities(Natural Sciences),2009,24(1):55-57. |
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| Authors: | WANG Wei ZHENG Gen-chang YAN Guang-zhao CHEN Bing-yan BAO Feng-li |
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| Affiliation: | WANG Wei, ZHENG Gen-chang, YAN Guang-zhao, CHEN Bing-yan,BAO Feng-li (1.Inner Mongolia University for Nationalities,Tongliao 028043,China; 2.Beijing Denong Need Co, Beijing 100101,China ) |
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| Abstract: | The factors influencing RAPD analysis, includingTaq polymerase, Mg2＋, dNTPs, primers, template DNA concentration, denaturing temperature in Dry peppers were studied. An optimal PCR system for RAPD in fr esh leaves of Dry peppers has been found：in 25 μ L reaction solution,contained Taq polymerasel.SU, Mg2＋2.5mmol/ L, dNTPs0.6mmol/L, primers0.8 μmol/L, template DNA6Ong. The amplification program was devised： initial denaturing at 94℃lot 4min;denaturing at 94℃ for lmin, annealing at 37℃ for l mins, extension at72℃ for 1.5min, 40 cycles;final extension at 72℃ for 5mins. |
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| Keywords: | RAPD-PCR |
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