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Número de catálogo | Descripción | N.º de reacciones |
---|---|---|
12596100 | Dye-free | 100 x 50 μL Reactions |
12596025 | Dye-free | 25 x 50 μL Reactions |
12597025 | Colored | 25 x 50 μL Reactions |
12597100 | Colored | 100 x 50 μL Reactions |
12596500 | Dye-free | 500 x 50 μL Reactions |
12597500 | Colored | 500 x 50 μL Reactions |
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.
Yes, reaction setup with SuperScript IV UniPrime One-Step RT-PCR System can be performed at room temperature. Moreover, the innovative two-phase hot start mechanism allows preassembled reactions to remain stable at room temperature for up to 24 hrs for targets up to 3 kb and for up to 4 hrs for targets >3 kb.
Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.
SuperScript IV UniPrime One-Step RT-PCR System includes 2X UniPrime RT-PCR master mix containing modified Platinum SuperFi II DNA Polymerase, which provides high specificity and high yields, and is ideally suited for applications that require sequence accuracy. On the contrary to common proofreading enzymes, this polymerase is compatible with dUTP.
Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.
Red and blue tracking dyes included in the colored format of SuperScript IV UniPrime One-Step RT-PCR System do not interfere with electrophoresis in E-Gel agarose gels. Please note that the reaction product obtained with both the colored and dye-free format should be diluted 2- to 30-fold for optimal separation using E-Gel agarose gels.
Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.
The two-phase hot-start mechanism, utilized in the SuperScript IV One-Step RT-PCR System and SuperScript IV UniPrime One-Step RT-PCR System, ensures sequential activation of RT and PCR enzymes in the one-step RT-PCR workflow. At ambient temperature, SuperScript IV RT is maintained inactive with a heat-sensitive RT-blocker. During the first hot-start activation phase at approximately 45 degrees C, the RT-blocker is released and the first-strand cDNA synthesis is initiated. During the second activation phase, the reaction is heated to 98 degrees C to activate DNA Polymerase and simultaneously inactivate SuperScript IV RT. This mechanism separates the RT and PCR enzymes' activities, delivering the highest RT-PCR specificity and yield.
Good laboratory practices are important for long fragment, one-step RT-PCR. These include using high-quality templates (pure, fresh, and intact) and fresh primer solutions. Optimization steps to consider include use of longer extension times and increasing template amounts. Learn more about RT-PCR reaction optimization and setup by visiting our reverse transcription educational resources (https://www.thermofisher.com/us/en/home/life-science/cloning/cloning-learning-center/invitrogen-school-of-molecular-biology/rt-education.html).
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