Every single pipetting step and each tube that has to be opened increases the risk of contamination. So far, reverse transcription involved the risk of contaminating the valuable sample by many pipetting steps. With our new all-in-one enzyme HotScriptase RT Polymerase > the risk of contamination is clearly minimized with the zero-step protocol.
Moreover, not to mention the considerable time saving.
In the two-step method, the lid of the reaction tube has to be opened after the cDNA synthesis, in order to convert it into the PCR reaction.
The one-step method has the disadvantage that a plurality of reagents have to be added.
Our HotScriptase RT Polymerase > provides a novel approach.
Reverse transcription and DNA amplification work simultaneously during a standard PCR. A "zero-step" RT-PCR is performed, which requires no further intermediate steps. Using appropriate Probes or Primers, quantitation of the original RNA content can be accomplished directly. There is no further pipetting or an additional program necessary. Thus, the risk of contamination is reduced considerably.
This risk can be further reduced by the use of HotScriptase RT Mastermix >. It contains all reagents except of template and primers.