Product category:
Polymerase chain reaction (PCR)
News Release from: Bio-Rad Laboratories | Subject: iScript
Edited by the Laboratorytalk Editorial
Team on 23 November 2004
Kits deliver quantitative RT-PCR in one
step
Couple the reverse transcriptase and real-time PCR steps of a quantitative RT-PCR reaction, by blending the components required for both steps in a single supermix
The new iScript One-Step quantitative RT-PCR kits couple the reverse transcriptase and real-time PCR steps of a quantitative RT-PCR reaction, by blending the components required for both steps in a single supermix Advantages of the new iScript one-step RT-PCR kits include:
This article was originally published on Laboratorytalk on 19 Feb 2002 at 8.00am (UK)
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Compatibility with all real-time detection chemistries - the iScript one-step RT-PCR kit with SYBR green is optimised for single-colour detection, while the iScript one-step RT-PCR kit for probes is optimised for probe hybridisation and hydrolysis in singleplex or multiplex RT-PCR.
The kits allow highly sensitive detection on a wide range of thermal cyclers.
cDNA synthesis and PCR amplification are conveniently carried out in the same tube, using the powerful combination of iScript RNase H+ reverse transcriptase and iTaq hot-start DNA polymerase.
The choice of reverse transcriptase can play a major role in the efficiency of the first-strand cDNA synthesis reaction.
Several attributes of the iScript reverse transcriptase allow it to effectively transcribe high yields of cDNA for PCR.
One of the key attributes of the iScript reverse transcriptase is that it is RNase H+.
RNase H+ reverse transcriptases degrade RNA molecules present in RNA-DNA hybrids, ensuring a clean pool of cDNA for PCR.
These reverse transcriptases increase the sensitivity of quantitative RT-PCR detection and analysis.
Thus, the sensitivity of both iScript kits exceeds that of other kits on the market, generating linear results down to 100fg of input, says Bio-Rad.
The iScript kit with SYBR green is one of the few kits that effectively minimises the incidence of primer-dimers, a common pitfall of the one-step RT-PCR procedure.
Primer-dimers can compete with the RT-PCR reaction being studied, and can lead to inaccurate results.
This kit contains a balanced combination of cations to prevent primer-dimer formation in a SYBR green background.
The iScript one-step RT-PCR kit for probes has been tested in a multiplex setting using two targets.
This is sasid to be the only kit that has demonstrated duplex reaction results in a one-step quantitative RT-PCR format.
The ability to duplex successfully is largely dependent on input template concentrations; both templates should be present in roughly equal amounts.
The iScript one-step RT-PCR kits are linear over the broadest dynamic range and are capable of detecting even trace amounts of input RNA.
In just one easy step, these kits provide a convenient and sensitive means of performing quantitative RT-PCR. Request a free brochure from Bio-Rad Laboratories ...
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