SuperScript IV UniPrime One-Step RT-PCR System

The Invitrogen SuperScript IV UniPrime One-Step RT-PCR System offers a convenient one-step process  for performing fast cDNA synthesis and PCR amplification in a single tube. It combines high-processivity SuperScript IV Reverse Transcriptase (RT) with a novel UniPrime RT-PCR master mix for easier use and unmatched  performance compared to other one-step RT-PCR kits.

Highlights
• Simplified workflow—universal primer annealing at 60°C decreases optimization time
• Easy pipetting—colored buffers for visual tracking of reaction setup and direct sample loading on gels
• Suitable for automation—two-phase hot-start activation mechanism for room temperature setup and benchtop stability for up to 24 hours
• Superior performance—high yields and reliable target detection even with challenging RNA samples

Combination of superior enzymes
The SuperScript IV UniPrime One-Step RT-PCR System utilizes a combination of superior enzymes. The SuperScript IV Reverse Transcriptase mix helps produce an efficient reverse transcription step through its high thermostability, processivity, and ability to synthesize cDNA from variety of RNA samples, while its high sensitivity helps ensure reliable target detection even when RNA input is limited (down to 0.01 pg RNA).

The 2X UniPrime RT-PCR master mix contains modified Platinum SuperFi II DNA Polymerase, which delivers high specificity and high yields and is ideally suited for applications that require sequence accuracy. This novel master mix formulation enables universal primer annealing at 60°C for most primer pairs designed following the general design rules and helps minimize optimization steps, save time, and avoid mistakes in RT-PCR runs.

Two-phase hot start
The innovative two-phase hot-start mechanism helps deliver high specificity and yield in one-step RT-PCR through temporal separation of the activities of the reverse transcriptase and DNA polymerase. This not only allows reaction setup at room temperature, but also helps ensure the stability of assembled reactions for up to 24 hours for targets up to 3 kb and for up to 4h for targets >3 kb.

Colored format
The SuperScript IV UniPrime One-Step RT-PCR System is availabe in a standard dye-free format or colored format containing tracking dyes. When the red tracking dye in the SuperScript IV RT mix and the blue tracking dye in the UniPrime RT-PCR master mix are combined during reaction setup, the final reaction mix turns to purple. This allows for simple visual tracking of reaction setup and helps prevent pipetting errors. The final reaction product can also be directly loaded on agarose gels for a more streamlined workflow.