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ABRF Study to Assess Priming Strategies for cDNA Synthesis in Real-Time qPCR


The Association of Biomolecular Resource Facilities' Nucleic Acid Research Group is seeking participants to help evaluate different priming strategies for cDNA synthesis in real-time quantitative PCR.

NARG said in a statement that the study is an opportunity for researchers to anonymously test their skills in performing cDNA synthesis using different primers, primer combinations, and no primer.

NARG said that has already tested 25 different priming strategies and "narrowed the primers in this follow-up study to those that performed the best in real-time RT-qPCR."

The primers were selected after testing using three transcripts that are expressed at different levels: beta-actin, beta-glucoronidase, and TATA binding protein.

NARG said that the study also aims to evaluate the impact of RNA integrity on cDNA synthesis.

Participants will receive two total RNA templates of varying quality, primers for the RT reactions, and instructions.

"Each participating laboratory will run the RT reaction using their chemistry of choice under conditions appropriate for that chemistry," NARG said. Each group will then return the tubes and plates containing the cDNA to a NARG member laboratory, "and fill out an online questionnaire detailing how the RT reaction was performed." NARG will perform the real-time PCR "in a single lab on a single instrument using the same PCR chemistry to minimize any variation introduced from the qPCR step."

The results of the study will be presented at the ABRF 2010 meeting, scheduled for March 20-23 in Sacramento, Calif. The results will also be posted on the ABRF website following the meeting and will be published.

Further information is available here.

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