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+Pwo DNA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Taq DNA聚合酶扩增的PCR产物主要得到3′端突出1个A(占67.3%,35/52);在Taq+Pwo DNA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.表明PCR扩增产物的末端是复杂多样的.
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.
夏庆杰,张思仲,武辉,肖翠英. PCR产物末端性质的分析研究[J].生物化学与生物物理进展,2000,27(2):215-217
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