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
