qScript XLT One-Step RT-PCR Kit

The qScript XLT One-Step RT-PCR Kit is a convenient and highly sensitive system for amplification of RNA templates up to 2 kb. cDNA synthesis and PCR amplification are carried out in the same tube without opening between procedures. This system has been optimized to deliver maximum RT-PCR efficiency, sensitivity, and specificity.qScript XLT is an engineered M-MLV reverse transcriptase with reduced RNase H activity and improved activity and stability at higher temperatures. The use of higher temperatures (48 to 55°C) for the cDNA synthesis step of one-step RT-PCR provides higher specificity for primer annealing and disruption of RNA secondary structure that can interfere with cDNA synthesis. The enzyme is supplied as a mixture with ribonuclease inhibitor protein to protect the integrity of RNA templates in crude lysates or samples where RNase contamination may limit assay sensitivity.A key component of the system is the One-Step ToughMix®. This master mix is highly resistant to PCR inhibitors and contains an ultra pure, hotstart, highly processive thermostable DNA polymerase that is blended with a proof-reading (3′-exonuclease) polymerase for improved PCR fidelity and fragment lengths. High-avidity monoclonal antibodies provide an extremely stringent automatic hot-start that minimizes the potential for primer-dimer and other non-specific PCR artifacts without compromising polymerase activities. Highly specific amplification is crucial to successful RT-PCR as non-specific product(s) can compete with amplification of the target sequence and impair PCR efficiency. The proprietary reaction buffer has been specifically formulated to maximize activities of both the reverse transcriptase and thermostable DNA polymerase while minimizing the potential for primer-dimer and other non-specific PCR artifacts.GelTrack® Loading Dye is a mixture of blue and yellow electrophoresis-tracking dyes that migrate at approximately 4kb and 50 bp. This optional component simplifies post PCR analysis, allowing direct loading of RT-PCR product on agarose gels following amplification. The GelTrack Loading Dye solution is not included with the sample kit.

Contents

• qScript XLT One-Step Reverse Transcriptase: 25X concentrated mixture of qScript XLT reverse transcriptase and recombinant ribonuclease inhibitor protein, 1 x 200 µL
• One-Step ToughMix: 2X concentrated reaction buffer containing dNTPs, magnesium chloride, hot-start DNA polymerase, and stabilizers, 2 x 1.25 mL
• GelTrack Loading Dye: 50X concentrated mixture of RT-PCR compatible, blue and yellow electrophoresistracking dyes, 1 x 0.4 mL
• Nuclease-free water 2 x 1.5 mL

Storage conditions

Kit components are stable for one year when stored in a constant temperature freezer at -20ºC. For convenience, the One-Step ToughMix may be stored unfrozen at +2 to +8°C for up to 6 months. Repeated freezing and thawing of the ToughMix will not affect product performance.

Precautions and Disclaimer

For research use only. Not intended for any animal or human therapeutic or diagnostic use.
Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

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