qScript™ One-Step qRT-PCR Kit

The qScript™One-Step qRT-PCR Kit is a convenient and highly sensitive solution for real-time quantitative PCR (qRT-PCR) of RNA templates using hybridization probe detection chemistries such as TaqMan® 5′-hydrolysis probes or molecular beacons on real-time quantitative PCR systems that do not require an internal reference dye. cDNA synthesis and PCR amplification are carried out in the same tube without opening between procedures. The system has been optimized to deliver maximum RT PCR efficiency, sensitivity, and specificity, enabling unbiased co-amplification of low copy transcripts in the presence of higher copy reference genes. The proprietary reaction buffer has been specifically formulated to maximize activities of both reverse transcriptase and Taq DNA polymerase while minimizing the potential for primer-dimer and other non-specific PCR artifacts. 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 this kit is AccuStart™ Taq DNA polymerase which contains monoclonal antibodies that bind to the polymerase and keep it inactive prior to the initial PCR denaturation step. Upon heat activation at 95°C, the antibodies denature irreversibly, releasing fully active, unmodified Taq DNA polymerase.

 

Features & Benefits

• Convenient single-tube reaction setup and streamlined protocol
• High sensitivity across a broad range of RNA template concentrations
• Duplex-optimized formulation – eliminates RT-PCR bias associated with high-copy reference genes, no primer optimization required

 Real-Time PCR Instrument Compatibility

• Roche LightCycler™ 480
• Qiagen/Corbett Rotor-Gene® Q
• Qiagen/Corbett Rotor-Gene® 6000
• Qiagen/Corbett Rotor-Gene® 3000
• Eppendorf Mastercycler® ep realplex
• Eppendorf Mastercycler® ep realplex2 s
• Cepheid SmartCycler®
• Bio-Rad CFX96™
• Bio-Rad CFX384™
• Bio-Rad/MJ Opticon®
• Bio-Rad/MJ Opticon 2
• Bio-Rad/MJ Chromo4™
• Bio-Rad MiniOpticon™
• Illumina Eco qPCR
• Thermo Scientific PikoReal Cycler
• Analytik Jena qTower
• BioRad iCycler iQ™
• BioRad MyiQ™
• BioRad iQ™5

Contents

• qScript One-Step Reverse Transcriptase – Optimized 50X formulation of recombinant MMLV reverse transcriptase for one-step RT-PCR.
• One-Step Master Mix (2X) – 2X reaction buffer containing dNTPs, magnesium chloride, AccuStart Taq DNA polymerase, and stabilizers
• Nuclease-free water

Storage conditions

Kit components are stable for one year when stored in a constant temperature freezer at -20°C. For convenience, the One-Step Master Mix may be stored unfrozen at +2 to +8°C for up to 6 months. Repeated freezing and thawing of the reaction mix is not recommended.

Resources

• 95057 qScript One-Step qRT-PCR kit Protocol
• 95057 qScript One-Step qRT-PCR kit MSDS

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