枸杞；_基因组DNA；_RAPD；_指纹图谱_

枸杞基因组DNA提取及指纹图谱分析

http://www.100md.com 《时珍国医国药》 2007年第1期

枸杞；,基因组DNA；,RAPD；,指纹图谱,,枸杞；,基因组DNA；,RAPD；,指纹图谱,1材料与方法,2结果,3讨论,参考文献：

     摘要：目的建立一种适于RAPD分析的快速提取枸杞叶片基因组DNA的方法，并利用RAPD技术研究枸杞属植物基因组DNA指纹图谱。方法采用改进的CTAB法提取枸杞冷藏幼叶的基因组DNA，利用RAPD技术对其进行指纹图谱分析。结果改进的CTAB法提取的10份枸杞材料基因组DNA均适于RAPD分析；可提供清晰的RAPD指纹图谱，利用筛选出的2个随机引物对供试材料DNA进行随机扩增，共得到56条带，其中多态性条带44条，多态性百分率为78.6%。结论表明改进的CTAB法提取的枸杞基因组DNA不需经氯化铯梯度离心或柱层析纯化，可以直接用于RAPD分析；RAPD对枸杞属植物进行指纹图谱分析是可行的。

    关键词：枸杞；基因组DNA； RAPD；指纹图谱

    Extraction of Genomic DNA from Lycium bararum and Its Fingerprint Analysis

    YAN Fengkun， XU Xing*， WEI Yuqing， YONG Lihua， ZHENG Guobao

    (Life Science School of Ningxia University, Yinchuan 750021, China； Key Lab of Agricultural Bio-technology of Ningxia, Ningxia Academy of Agricultural and Forestryial Sciences, Yinchuan 750002, China； Agricultural College of Ningxia University, Yinchuan 750021, China)

    Abstract：ObjectiveTo develop valid method to extract DNA from Lycium bararum for RAPD analysis and research its DNA fingerprint by using RAPD technology. MethodsThe extracts of high quality genomic DNA was extracted from the frozen leaves of Lycium bararum by modified CTAB method and its fingerprint was analyzed by RAPD. ResultsThe clear fingerprints of the extracts of genomic DNA could be got by RAPD. Two RAPD primers were applied to do random amplification. A total of 56 DNA bands was detected, 44 among which were polymorphic, the average rate of polymorphic bands was 78.6%.ConclusionThe modified CTAB method was valid,quick and didn＇t need phenol extraction. This isolated DNA could be used directly for RAPD analysis without sedimentation in cesium chloride gradient or column chromatography.The RAPD markers can be used for the studies of fingerprint of Lycium bararum sensitively. ......

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