Poly(A) cDNA-specific (PACS) RT-PCR: A quantitative method for the measurement of any poly(A)-containing mRNA not affected by contaminating genomic DNA
Poly(A) cDNA-specific (PACS) RT-PCR: A quantitative method for the measurement of any poly(A)-containing mRNA not affected by contaminating genomic DNA
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DOI：
10.2144/00294st01
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发表时间：
2000-10-01
期刊：
影响因子：
2.7
通讯作者：
Nepluev, I
中科院分区：
文献类型：
--
作者：
Folz, RJ;Nepluev, I
我们提出了一种简单而有效的RT-PCR方法来检测和定量任何含有聚(A)的mRNA，该方法不受污染的基因组DNA的影响，也不依赖于详尽的DNA酶消化方案。这里描述的技术需要使用设计为包含6-8bpc DNA特异序列的反义引物和位于5‘端的另外17个TS来利用Poly(A)尾。可以使用不需要通过内含子DNA序列分离的第二个cDNA特异性正义引物。
We present a simple and efficient RT-PCR method for the detection and quantitation of any poly(A)-containing mRNA that is not affected by contaminating genomic DNA and does not rely on exhaustive DNase digestion protocols. The technique described here requires the use of an antisense primer designed to contain 6-8 bp cDNA-specific sequence and an additional 17 Ts located on the 5' end to take advantage of the poly(A) tail. A second cDNA-specific sense primer can be used that does not need to be separated by intronic DNA sequence.