qScript™ One-Step qRT-PCR Kit, Low ROX™

The qScript™One-Step qRT-PCR Kit, Low ROX™ 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 Applied Biosystems 7500 or Stratagene MX series of real-time PCR systems. 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

• Applied Biosystems 7500
• Applied Biosystems 7500 Fast
• Stratagene Mx3000P®
• Stratagene Mx3005P™
• Stratagene Mx4000™
• Applied Biosystems ViiA 7
• Applied Biosystems QuantStudio™ 12K Flex

Contents

• qScript One-Step Reverse Transcriptase – Optimized 50X formulation of recombinant MMLV reverse transcriptase for one-step RT-PCR.
• One-Step Master Mix, Low ROX (2X) – 2X reaction buffer containing dNTPs, magnesium chloride, AccuStart Taq DNA polymerase, ROX Reference Dye (for 580-585 nm excitation), and stabilizers
• Nuclease-free water

Storage conditions

Kit components are stable for one year when stored in a constant temperature freezer at -20°C protected from light. For convenience, the One-Step Master Mix, Low ROX 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

• 95059 qScript One-Step qRT-PCR kit, Low ROX Protocol
• 95059 qScript One-Step qRT-PCR kit, Low ROX MSDS

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