Adaptor-tagged competitive PCR: A novel method for measuring relative gene expression
Adaptor-tagged competitive PCR: A novel method for measuring relative gene expression
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DOI：
10.1093/nar/25.22.4694
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发表时间：
1997-11-15
影响因子：
14.9
通讯作者：
Kato, K
中科院分区：
文献类型：
--
作者：
Kato, K
描述了一种简单可靠的基于PCR的基因表达定量方法。在用限制性内切酶消化双链cDNA之后，将接头连接到来自第一RNA样品的cDNA，并且将另一接头连接到第二RNA样品。将等量的连接样品混合，并通过接头引物和对目的基因特异性的引物扩增。来自两种来源的产物大小不同，并且可以通过变性聚丙烯酰胺凝胶电泳分离。两种产物的比率揭示了基因表达的相对水平。由于该技术避免了构建内标的需要，它对于分析许多不同的基因转录物特别有用。
A simple and reliable PCR-based method to quantitate gene expression is described, Following the digestion of double-stranded cDNA by a restriction enzyme, an adaptor is ligated to a cDNA from a first RNA sample, and another adaptor to a second RNA sample, The two adaptors share a common sequence at the outer region, but differ in size. Equal amounts of the ligated samples are mixed, and amplified by an adaptor-primer and a primer specific to the gene of interest, Products derived from the two sources differ in size, and can be separated by denaturing polyacrylamide gel electrophoresis, The ratio of the two products reveals the relative level of gene expression, Since the technique avoids the need to construct internal standards, it is especially useful for the analysis of many different gene transcripts.