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RT-PCR: minus-sense RNA detection

Rod Pennington OKIEROD at UKCC.UKY.EDU
Tue Jan 17 08:29:01 EST 1995


Greetings from the Bluegrass:

Occasionally I hear or read of RT-PCR detection of minus-sense
viral RNA based on using RT primers specific for the minus-sense
RNA (thus, any cDNA available for PCR amplification is a supposedly
a product of reverse transcription of minus-sense RNA).  However,
often no acknowledgement is made of the possibility, or probability,
that the cDNA produced in the RT reaction could be the result of
self-priming of the plus-sense RNA, ie, there really is no strand
specificity in the RT-PCR reaction.  This topic came up at least
once before here on the list and useful suggestions for eliminating
self-priming were mentioned.

My question: Does anybody have a good reference which specifically
addresses the questions of RNA self-priming or minus-sense RNA
detection by RT-PCR?

Thanks

Rod Pennington
Department of Plant Pathology
University of Kentucky
okierod at ukcc.uky.edu



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