香蕉RAPD反应体系的建立

中国农学通报 ›› 2009, Vol. 25 ›› Issue (16): 51-55.

香蕉RAPD反应体系的建立

刘金福,潘东明,代立春,庄西卿,李英豪,赖钟雄,吴少华

收稿日期:2009-02-24 修回日期:2009-04-30 出版日期:2009-08-20 发布日期:2009-08-20

The Establishment of RAPD Reaction Program of Fujian Banana

Liu Jinfu,, Pan Dongming,, Dai Lichun, Chen Qingxi, Zhuang Xiqing，Li Yinghao

Received:2009-02-24 Revised:2009-04-30 Online:2009-08-20 Published:2009-08-20

摘要/Abstract

摘要： 利用改良的CTAB方法从香蕉叶片中提取高质量的DNA。在参考一般RAPD分析反应程序的基础上，经过反复试验，确定适合香蕉PCR扩增程序为：94℃预变性 5min；94℃变性 1min；38℃退火 1min；72℃延伸 2min；变性ó延伸，循环45次；最后72℃延伸 2min。PCR扩增的体系（总体积25μL）为：模板DNA 20ng，dNTP 200μmol/L，10×PCR Buffer2.5μL，引物0.20μmol/L，Taq酶0.75U，ddH2O17.85μL。

Abstract: Using the improved method of CTAB，high purity DNA was obtained from banana leaves．Based on the common RAPD reaction program and adjusting experiments，the optimal amplification program was described as follows：94℃ for 5min；45 cycles at 94℃ for 1 min，38℃ for 1 min and 72℃ for 2 min；72℃ for 2min at last. PCR system（25μL total volumes）contains：20ng of genomic DNA，200200μmol/L of dNTP，2.5μL of 10×PCR Buffer，0.20μmol/L of primer，0.75U Taq DNA polymerase，17.85μL of ddH2O． Key words：banana；Template DNA；RAPD

刘金福,潘东明,代立春,庄西卿,李英豪,赖钟雄,吴少华. 香蕉RAPD反应体系的建立[J]. 中国农学通报, 2009, 25(16): 51-55.

Liu Jinfu,, Pan Dongming,, Dai Lichun, Chen Qingxi, Zhuang Xiqing，Li Yinghao. The Establishment of RAPD Reaction Program of Fujian Banana[J]. Chinese Agricultural Science Bulletin, 2009, 25(16): 51-55.