逆转录酶
Different types of reverse transcriptase polymerases are available to suit different needs and reaction conditions. Reverse transcriptase enzymes commonly used include AMV RT, SuperScript III, ImProm-II, Omniscript, Sensiscript and Tth RT. At the end of the RT step the polymerase enzyme is heatactivated. It could also be that the reverse transcriptase polymerase and DNA polymerase is one and the same enzyme and that the enzyme only requires a DNA polymerase activation step after the RT step. An example of such an enzyme is Tth polymerase. This enzyme has both RNA-dependent reverse transcriptase and DNA-dependent polymerase activity. However, the active center of the DNA polymerase is covered by dedicated oligonucleotides, called aptamers. At temperatures below the optimal reaction temperature of the DNA-dependent polymerase component of Tth remains covered by the aptamers. At these temperatures the Tth enzyme only synthesizes a DNA copy of the RNA template. Once the reaction temperature is raised to 95 °C, the aptamers are removed and the DNA-dependent polymerase component will start to amplify the target sequence.