Abstract
Reverse transcription of RNA followed by the polymerase chain reaction (RT-PCR or RNA-PCR) is an extraordinarily sensitive method to detect as few as 1–100 copies of a specific RNA (1-3). However, we and others have found that false positives caused by contamination with minute quantities of DNA (i.e., cDNAs, plasmid DNAs, genomic DNA, or PCR carryover) is a major shortcoming of the method even when meticulous laboratory technique is employed (4-6). RNA template-specific PCR (RS-PCR) is a modification of conventional RNA-PCR in which RNA is reverse-transcribed with a primer that contains at its 5′ end a unique nucleotide sequence that may then be exploited in the PCR to amplify preferentially the RNA-derived sequence. RS-PCR retains full sensitivity, but reduces dramatically the frequency of false positives (7-9).
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© 1993 Humana Press Inc., Totowa, NJ
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Shuldiner, A.R., Perfetti, R., Roth, J. (1993). RNA Template-Specific Polymerase Chain Reaction (RS-PCR). In: White, B.A. (eds) PCR Protocols. Methods in Molecular Biology, vol 15. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-244-2:169
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DOI: https://doi.org/10.1385/0-89603-244-2:169
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-244-6
Online ISBN: 978-1-59259-502-0
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