Shuldiner A R, Nirula A, Roth J
Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.
Gene. 1990 Jul 2;91(1):139-42. doi: 10.1016/0378-1119(90)90176-r.
We report a novel modification of the reverse transcription-polymerase chain reaction method that we have dubbed RNA template-specific PCR (RS-PCR). With this approach, the 5' end of the first strand is tagged with a unique nucleotide sequence during reverse transcription which may then be exploited to amplify preferentially RNA-derived sequences. In our hands, RS-PCR greatly reduces the frequency of false positives and virtually eliminates carryover contamination from DNA fragments amplified in previous experiments.
我们报告了一种对逆转录-聚合酶链反应方法的新颖改进，我们将其命名为RNA模板特异性PCR（RS-PCR）。通过这种方法，在逆转录过程中，第一链的5'端会被标记上一个独特的核苷酸序列，随后可利用该序列优先扩增RNA衍生的序列。在我们的操作中，RS-PCR极大地降低了假阳性的频率，并且几乎消除了先前实验中扩增的DNA片段带来的残留污染。
