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
Folz, R.J.; Nepluev, I.
Biotechniques 29(4): 762 764
2000
ISSN/ISBN: 0736-6205 PMID: 11056805 Document Number: 518802
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.