RAPD扩增中商品酶体系对扩增产物的影响  

DISTINCT AMPLIFIED PRODUCTS BY DIFFERENT COMMERCIAL Taq DNA POLYMERASE IN RAPD

作　　者：何礼[1] 袁水全[2] 莫邦辉[2] 姚光贵 林宏辉[1] 陈放[1] 郭骁才[2] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]中国科学院成都生物研究所,成都610041 [3]成都市第一农业科学研究所,成都610072

出　　处：《应用与环境生物学报》2002年第3期308-313,共6页Chinese Journal of Applied and Environmental Biology

基　　金：四川省重点基础研究项目 ;成都市科技攻关重点项目 ;中国科学院知识创新工程项目;国家自然科学基金资助项目 (No.30 1 70 557)

摘　　要：随机选用了 10条RAPD引物 ,采用 3种商品TaqDNA聚合酶体系 ,以 5个不同物种的动植物总DNA为模板 ,进行RAPD扩增 .实验发现 ,在对同一模板、采用同一引物进行的RAPD扩增中 ,仅仅因为使用了不同的商品TaqDNA聚合酶 ,获得的扩增产物差别极大 ;不同商品来源的PCR反应缓冲体系 ,对扩增产物也有一定影响 ,但相对较小 .这说明 ,不同的商品酶体系对RAPD扩增产物影响很大 .因此 ,影响RAPD扩增产物稳定性的一个关键因素是商品TaqDNA聚合酶本身 ;研究中前后一致地使用同一商品TaqDNA聚合酶体系 ,是成功进行RAPD分析的基础 .图 3参The total DNAs from 3 species of plants and 2 species of animals were amplified by RAPD (random amplified polymorphic DNA) using 10 RAPD 10 nt primers and 3 different commercial Taq DNA polymerases, respectively. The results indicated that distinct amplified fragments were obtained in the same reaction of RAPD amplification by using the same plant or animal DNA as template and using the same RAPD primer, but the commercial Taq DNA polymerases used were different. Identical or a few different amplified fragments were obtained when the PCR 10× buffers from different commercial products were used in the same reaction of RAPD in which the same template DNA, primer and commercial Taq DNA polymerase were used. So the amplification of RAPD were influenced mostly by the commercial Taq DNA polymerases. A successful analysis of RAPD might rely on using the same commercial Taq DNA polymerase before chosen. Fig 3, Ref 19

关 键 词：RAPD扩增 商品酶体系 扩增产物 

分 类 号：Q503[生物学—生物化学]