DeFilippes F M
Laboratory of Viral Diseases NIAID, NIH, Bethesda, MD 20892.
Biotechniques. 1991 Jan;10(1):26, 28, 30.
Template DNA, which was added to a modified polymerase chain reaction mixture to simulate contamination, was inactivated by restriction enzyme digestion. After inactivation of the restriction enzymes, additional template, buffer and Taq polymerase were added to the reaction and the polymerase chain reaction proceeded. Results are presented using several different restriction enzymes with four base recognition sequences. The limitations of the method are discussed to indicate the tactics to be used for other DNA sequences.
将模板DNA添加到改良的聚合酶链反应混合物中以模拟污染,然后通过限制性酶切消化使其失活。在限制性酶失活后,向反应中加入额外的模板、缓冲液和Taq聚合酶,然后进行聚合酶链反应。使用几种具有四碱基识别序列的不同限制性酶展示结果。讨论了该方法的局限性,以表明针对其他DNA序列应采用的策略。