cDNA equalization for reverse transcription-polymerase chain reaction quantitation.
cDNA equalization for reverse transcription-polymerase chain reaction quantitation.
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用于逆转录聚合酶链式反应定量的 cDNA 均衡。
DOI：
10.1006/abio.1993.1044
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发表时间：
1993
影响因子：
2.9
通讯作者：
Larson,J
中科院分区：
文献类型：
--
作者：
Kolls,J;Deininger,P;Cohen,JC;Larson,J
逆转录与聚合酶链反应的结合已越来越多地用于研究基因表达。然而，大多数先前发表的定量技术受到精确的初始RNA定量的限制，并且不能很好地解释逆转录的相对效率。我们已经开发了一种技术，标记和定量的随机引物cDNA产物的逆转录反应。使用聚合酶链反应结合模板稀释或与内部竞争模板，我们表明，通过正常化cDNA输入到聚合酶链反应，我们得到基因表达的准确定量。我们称这个过程为逆转录-聚合酶链反应的cDNA均衡化。这种方法是理想的小临床样品，其中输入RNA的准确定量是困难的。
Reverse transcription coupled with the polymerase chain reaction has been increasingly utilized to study gene expression. However, most previously published quantitative techniques are limited by accurate initial RNA quantitation and do not account well for the relative efficiency of reverse transcription. We have developed a technique of labeling and quantitating the random-primed cDNA product of a reverse transcription reaction. Using the polymerase chain reaction in conjunction with template dilutions or with an internal competitive template, we show that by normalizing the cDNA input into the polymerase chain reaction, we get accurate quantitation of gene expression. We call this procedure cDNA equalization of reverse transcriptase-polymerase chain reaction. This method is ideal for small clinical samples where accurate quantitation of input RNA is difficult.