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

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