Kits offer a convenient and highly sensitive solution for real-time reverse transcriptase PCR (qRT-PCR) of RNA templates using hybridization probe detection chemistries such as TaqMan® 5'-hydrolysis probes or molecular beacons on PCR instrument platforms.
For Minor Groove Binder (MGB) modified probes, qScript™ One-Step fast MGB qRT-PCR kits are recommended. These kits offer a convenient and highly sensitive solution for qRT-PCR of RNA templates using TaqMan-MGB 5'-hydrolysis probe detection exclusively on qRT-PCR instrument platforms.
The system allows cDNA synthesis and PCR amplification to be carried out in the same tube without opening between procedures. It is ideal for highly sensitive quantification of RNA viruses or low abundance RNA targets, as wells as high throughput gene-expression studies. It has also been optimized to deliver maximum RT-PCR efficiency, sensitivity, and specificity in reduced reaction volumes and fast cycle times.
One-step fast master mixes are provided as a 4x concentrated solution to allow addition of higher amounts of RNA template and improved detection sensitivity with low concentration samples. The unique formulation maximizes the activities of both reverse transcriptase and Taq DNA polymerase while minimizing the potential for primer-dimer and other non-specific PCR artifacts. This enables unbiased co-amplification of low copy transcripts in the presence of higher copy reference genes in duplexed qRT-PCR applications.
Highly-specific amplification is crucial to successful qRT-PCR, as non-specific product(s) can compete for amplification of the target sequence and impair PCR efficiency. A key component of these kits is AccuStart™ Taq DNA polymerase with monoclonal antibodies that bind to the polymerase and keep it inactive during reaction assembly and the 50°C reverse transcription step. A brief 30-second heat activation step at 95°C irreversibly denatures the antibodies, releasing fully active, unmodified Taq DNA polymerase. Rapid recovery of fully active, unmodified Taq DNA polymerase is critical for efficient extension kinetics. Therefore, these kits afford greater reagent economy and laboratory throughput on conventional or rapid ramp rate qPCR systems.
Kits are available for all real-time PCR instrument platforms including those requiring normalization with ROX™ reference dye. Platforms not requiring passive reference dye include Roche LightCycler® 480, Bio-Rad® iQ™, MyiQ™, iQ™5, Opticon®, Chromo4™, Corbett Rotor-Gene®, Eppendorf® Mastercycler®, and Cepheid® SmartCycler®. Kits with Low ROX™ reference dye are for use with platforms including AB 7500 and Stratagene® Mx™. Kits with ROX™ reference dye are for use with platforms including AB 7000, 7300, 7700, 7900, and StepOne™.
Ordering information: If not using Minor Groove Binder (MGB) modified probes exclusively, qScript™ One-Step Fast qRT-PCR kits are recommended.
