Reverse Transcription and Polymerase

ThermaStop-RT - ThermaGenix Inc.

RNA oligonucleotides from cells and viruses can be amplified for detection using the methods of Reverse Transcription and Polymerase Chain Reaction (RT-PCR).  In a first step, a complementary strand of DNA (cDNA) is synthesized using a primer and reverse transcriptase.  The cDNA is then amplified using standard PCR techniques.  This can be accomplished by taking an aliquot from the reverse transcription reaction and adding a DNA-dependent DNA polymerase (e.g. Taq) and other reagents and then doing PCR.  That method is commonly called "Two-step RT-PCR", or "Two-tube RT-PCR".  Alternatively, all reagents, including both enzymes and gene-specific primers can be included in a single tube in order to do both reverse trancription and PCR without the need to open the tube between those reactions.  That method is commonly called "One-Step RT-PCR" or "One-Tube RT-PCR".  Both methods can include primers for a single target or for multiple targets (i.e. multiplex RT-PCR).