The qScript™ XLT One-Step RT-PCR Kit is a convenient and highly sensitive system for amplification of RNA templates up to 2 kb

• ToughMix reagent technology withstands PCR inhibitors
• Proof-reading polymerase supports high-fidelity downstream applications
• Temperature stabilized to support reaction assembly at ambient room temperatures
• Optional GelTrack dye streamlines workflow for gel electrophoresis

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 4 kb 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.

Kit components are stable for one year when stored in a constant temperature freezer at -20°C. 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.

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.