PCR产物末端性质的分析研究

Studies on Termini of PCR Products.

利用PCR、UT PCR、克隆及测序等技术 ,对强直性肌营养不良基因 (MT PK) 3′ 非翻译区分别用Taq ,Taq +PwoDNA聚合酶进行了扩增、克隆和测序 ,研究了PCR产物末端组成情况 ,并比较了上述两种DNA聚合酶对PCR产物末端的影响 .结果在用TaqDNA聚合酶扩增的PCR产物主要得到 3′端突出 1个A (占 6 7 3% ,35 /5 2 ) ;在Taq +PwoDNA聚合酶扩增的PCR产物末端中得到 3′端 +A的仅占 17 4% ,而 - 1的占 34 8% ,与前者显著不同 .

PCR products amplified with Taq and Taq+Pwo DNA polymerase were reamplified by using UT PCR and then cloned and sequenced. The termini of the original PCR products were analyzed . In the group of Taq amplification , the termini were chiefly sticky ones with 3′ protruding A(67 3%);the next majority of termini were blunt ones(26 9%).In the other group, the proportion of blunt ends was almost the same as that in the group of Taq amplification (26 1%). However, the 3′ protruding termini decreased(17 4%),instead of additional 3′ recessive ones(-1,-2,-3;57 0%). The results reveal that the ends of PCR products are complicated .