What is the right reverse transcription method for your qPCR experiment?--Taq Talk Episode 9

The video introduces the topic of reverse transcription (RT) methods for qPCR experiments and explains that two primary approaches exist: two-step and one-step methods.

Describe the two-step approach where RNA is first converted to first-strand cDNA in step one, which can be stored or immediately used for second-strand cDNA synthesis and real-time detection in step two.

Describe the one-step approach where reverse transcription and PCR amplification occur simultaneously in the same tube, combining both processes in a single reaction.

Explain when to use the two-step method: for analyzing multiple targets in separate reactions from the same cDNA synthesis, and when sample quantities are limited since cDNA can be stored for later use.

Explain when to use the one-step method: for faster workflows, high-throughput gene expression analysis, RNA virus detection, liquid-handling robotics to reduce contamination, and when rapid time-to-results is critical.

After choosing the RT approach, select additional parameters for your qPCR experiment including detection chemistry options like TaqMan or SYBR Green and appropriate reagent formulations.

Direct viewers to use the online reagent selection guide at thermofisher.com to find the best RT and qPCR detection method reagents for their specific experimental needs.