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  7. Invitrogen™ SuperScript IV UniPrime One-Step RT-PCR System

Invitrogen™ SuperScript IV UniPrime One-Step RT-PCR System

Product Code. 17843054
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Description:
Colored
Dye-free
No. of Reactions:
100 x 50 μL Reactions
25 x 50 μL Reactions
500 x 50 μL Reactions
Unit Size:
100 reactions
25 reactions
500 reactions
6 product options available for selection
Product selection table with 6 available options. Use arrow keys to navigate and Enter or Space to select.
Product Code. Description No. of Reactions unitSize
17843054 Dye-free 100 x 50 μL Reactions 100 reactions
17871544 Dye-free 25 x 50 μL Reactions 25 reactions
17881544 Colored 25 x 50 μL Reactions 25 reactions
17832994 Colored 100 x 50 μL Reactions 100 reactions
17804584 Dye-free 500 x 50 μL Reactions 500 reactions
17814604 Colored 500 x 50 μL Reactions 500 reactions
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Product Code. 17843054 Supplier Invitrogen™ Supplier No. 12596100

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1189.40 EUR valid until 2026-05-31
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Description

Invitrogen SuperScript IV UniPrime One-Step RT-PCR System is a convenient one-step formulation for performing fast cDNA synthesis and PCR amplification in a single tube.

Invitrogen SuperScript IV UniPrime One-Step RT-PCR System is a convenient one-step formulation for performing fast cDNA synthesis and PCR amplification in a single tube. It combines the high-processivity SuperScript IV Reverse Transcriptase (RT) with a novel UniPrime RT-PCR Master Mix to provide easier use and unmatched performance when compared to other one-step RT-PCR kits.

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.

TRUSTED_SUSTAINABILITY

Specifications

Content And Storage • SuperScript IV RT Mix (200 μL)
• 2X UniPrime RT-PCR Master Mix (2 x 1.25 mL)
• Nuclease-free water (2 x 1.25 mL)

Store at -25°C to -15°C.
Description Dye-free
Detection Method Gel Electrophoresis
Format 1-step RT-PCR Kit
GC-Rich PCR Performance High
PCR Method 1-step RT-PCR
Technique 1-step RT-PCR
Multiplex Capability Yes
Optimal Reaction Temperature 50°C to 65°C
Quantity 100 reactions
Reverse Transcriptase SuperScript IV
Hot Start Built-in Hot Start
No. of Reactions 100 x 50 μL Reactions
Reaction Format Master Mix
Reagent Type Reverse Transcription
Size (Final Product) Up to 13 kb
Starting Material RNA
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Frequently Asked Questions (FAQs)

Can I set up my reactions at room temperature using SuperScript IV UniPrime One-Step RT-PCR System?

Yes. Reaction setup with SuperScript IV UniPrime One-Step RT-PCR System can be performed at room temperature. Moreover, due to innovative two-phase hot start mechanism, preassembled reactions remain stable at room temperature for up to 24 hours for targets up to 3 kb and up to 4 h for targets >3 kb.

What DNA Polymerase is included in SuperScript IV UniPrime One-Step RT-PCR System?

SuperScript IV UniPrime One-Step RT-PCR System includes 2X UniPrime RT-PCR Master Mix containing modified Invitrogen Platinum SuperFi II DNA Polymerase, which offers high specificity, high yields, and is ideally suited for applications that require sequence accuracy. Contrary to common proofreading enzymes, this polymerase is compatible with dUTP.

Will the tracking dyes in SuperScript IV One-Step RT-PCR System Colored interfere with fragment analysis in E-Gel agarose gels?

Red and blue tracking dyes included in the colored format of SuperScript IV One-Step RT-PCR System do

Can I set up my reactions at room temperature using SuperScript IV UniPrime One-Step RT-PCR System?

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.
Which DNA polymerase is included in SuperScript IV UniPrime One-Step RT-PCR System?

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.
Do the tracking dyes in the colored format of SuperScript IV UniPrime One-Step RT-PCR System interfere with electrophoresis in E-Gel agarose gels?

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.
What is a two-phase, hot-start activation mechanism?

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.
I am having difficulties performing one-step RT-PCR with long targets. What are your recommendations?

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. Consider optimizing the 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).

For RT-PCR targets up to 13 kb, we recommend SuperScript IV UniPrime One-Step RT-PCR System (Cat. No. 12596025).
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