FAQ: How does One-Step RT-PCR kit protocol work for NEB #E5315?

The One-Step RT-PCR reactions allow both cDNA synthesis and PCR amplification in the same reaction buffer. A thermocycler is programmed so that cDNA synthesis takes place first (48°C for 15 minutes), then reverse transcriptase is inactivated while Hot Start Taq Polymerase is activated (94°C for 1 minute), followed by 40 cycles of PCR amplification.