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

R. J. Folz; I. Nepluev

doi:10.2144/00294st01

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

R. J. Folz, I. Nepluev

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10 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	762-768
Number of pages	7
Journal	BioTechniques
Volume	29
Issue number	4
DOIs	https://doi.org/10.2144/00294st01
State	Published - 2000

ASJC Scopus subject areas

Biotechnology
General Biochemistry, Genetics and Molecular Biology

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AU - Nepluev, I.

PY - 2000

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AB - 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.

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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

Abstract

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