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.
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.
Folz
R J
RJ
Duke University Medical Center, Durham, NC 27710, USA. rodney.folz@duke.edu
Nepluev
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I
D000818:Animals; D018076:DNA, Complementary / Q000235:genetics*; D051379:Mice; D011061:Poly A / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D013482:Superoxide Dismutase / Q000235:genetics
D018076:DNA, Complementary; D012333:RNA, Messenger; D011061:Poly A; D013482:Superoxide Dismutase