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SuperScript™ III One-Step RT-PCR System with Platinum™ Taq High Fidelity DNA Polymerase (Invitrogen™)

The SuperScript III One-Step RT-PCR System with Platinum Taq High Fidelity is designed for convenient end-point detection and analysis of RNA molecules by one-step RT-PCR. This system consists of two major components: SuperScript III RT/ Platinum Taq High Fidelity Enzyme Mix and 2X Reaction Mix. The enzyme mix combines SuperScript III Reverse Transcriptase and Platinum Taq DNA Polymerase High Fidelity, which is a an enzyme mixture composed of recombinant Taq DNA polymerase, Pyrococcus species GB–D polymerase, and Platinum Taq antibodies, which block polymerase activity at ambient temperatures enabling hot start PCR. The 2X Reaction Mix consists of a proprietary buffer system optimized for reverse transcription and PCR amplification, Mg2+ optimized for universal use, deoxyribonucleotide triphosphates, and stabilizers. All components for cDNA synthesis and PCR are combined in a single tube with gene-specific primers and target RNA. Reverse transcription automatically follows PCR cycling without additional steps. The system can detect a wide range of RNA targets (up to 10 kb in length) at variable concentrations (1 pg to 1 µg of total RNA).

Note: For superior one-step RT-PCR performance, we recommend the SuperScript IV One-Step RT-PCR System or SuperScript IV One-Step RT-PCR System with ezDNase. The SuperScript IV One-Step RT-PCR System combines the high processivity of SuperScript IV Reverse Transcriptase with the high-fidelity of Platinum SuperFi DNA Polymerase to provide unmatched product yields, specificity, and sensitivity in less time and for a broad target range, even with suboptimally pure RNA samples.

SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase (Invitrogen™)

The SuperScript® III One-Step RT-PCR System with Platinum® Taq DNA Polymerase is designed for the sensitive, reproducible, end-point detection and analysis of RNA molecules by RT-PCR. Using this convenient one-step formulation, you can perform both cDNA synthesis and PCR amplification in a single tube using gene-specific primers, and target RNAs from either total RNA or mRNA. The system uses a mixture of SuperScript® III Reverse Transcriptase and Platinum® Taq DNA polymerase in an optimized reaction buffer, and it can detect a wide range of RNA targets, from 200 bp to 4.5 kb. The amount of starting material can range from 0.01 pg to 1 µg of total RNA. Key advantages of this kit:

Sensitive—routine detection down to 0.01 pg total RNA (see figure)
Convenience—one-step format for speed, convenience, and less reaction-to-reaction variability
Specificity—uses SuperScript® III RT for cDNA synthesis up to 55°C, for more specific priming with gene specific primers (see figure)
Amplicon size—compatible detection of targets up to 4.5 kb in length for greater flexibility

SuperScript® III Reverse Transcriptase
SuperScript® III Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme can synthesize cDNA at a temperature range of 45–60°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. Because SuperScript® III RT is not significantly inhibited by ribosomal and transfer RNA, it can be used to synthesize cDNA from total RNA.

Platinum® Taq DNA polymerase
Platinum® Taq DNA polymerase is recombinant Taq DNA polymerase complexed with a proprietary antibody that blocks polymerase activity at ambient temperatures. Activity is restored after the denaturation step in PCR cycling at 94°C, providing an automatic "hot start" in PCR for increased sensitivity, specificity, and yield.

High-Capacity RNA-to-cDNA™ Kit (Applied Biosystems™)

The High Capacity RNA-to-cDNA Kit is a streamlined reverse transcription kit designed for optimum performance with TaqMan Gene Expression Master Mix, Power SYBR Green PCR Master Mix, and other PCR enzymes.

Features of this kit include:
Fast—short reaction time (typically 30–60 min)
Convenient—easy workflow with few pipetting steps
Reliable—reverse transcription of both abundant and limited targets
Flexible—optimized 2-step protocol, enabling multiple PCR reactions from a single reverse transcription reaction

Streamline your cDNA synthesis while maintaining detection sensitivity
The components of the two-tube kit work together to provide sensitive and specific reverse transcription across a broad range of template amounts. Amplification of a dilution series of the RNA concentration reference standard demonstrates exceptional reverse transcription and PCR efficiency across 11 orders of magnitude. This wide dynamic range allows one set of reaction conditions to detect transcripts from highly active as well as weakly expressed genes. The two-tube formulation reduces reaction time and requires fewer pipetting steps than with the High Capacity cDNA Reverse Transcription Kit, while maintaining high performance.

SuperScript™ IV One-Step RT-PCR System (Invitrogen™)

The Invitrogen SuperScript IV One-Step RT-PCR System combines the high processivity of SuperScript IV Reverse Transcriptase (RT) with the high-fidelity of Platinum SuperFi DNA Polymerase to provide unmatched one-step RT-PCR performance. With this convenient one-step formulation plus gene-specific primers, you can perform both cDNA synthesis from total RNA or mRNA and PCR amplification in a single tube.

Features of the SuperScript IV One-Step RT-PCR System include:
• Superior performance with sensitivity down to 0.01 pg RNA, target length up to 13.8 kb, and the fastest one-step RT-PCR protocol
• Reliable target detection even with suboptimally pure RNA samples
• Two-phase hot-start activation mechanism for high sensitivity, specificity, and room temperature setup

SuperScript IV RT is a proprietary MMLV RT mutant with increased processivity, thermostability, and inhibitor resistance, as well as reduced RNase H activity, for sensitive and efficient full-length cDNA synthesis. Platinum SuperFi DNA Polymerase is a proofreading DNA polymerase that combines superior fidelity with trusted Platinum hot-start technology for the highest success in PCR. The combination of these two engineered enzymes produces enhanced RT-PCR yields and fidelity, as well as amplification of longer templates compared to other one-step RT-PCR kits.

For even better performance, we offer SuperScript IV One-Step RT-PCR System with ezDNase. The ezDNase further enhances the one-step RT-PCR workflow through an extremely simplified genomic DNA removal step that ensures the highest accuracy and confidence in one-step RT-PCR results.

SuperScript™ IV First-Strand Synthesis System with ezDNase™ Enzyme (Invitrogen™)

The SuperScript IV First-Strand Synthesis System with ezDNase Enzyme for RT-PCR is optimized for synthesis of first-strand cDNA from purified poly(A)+ or total RNA. The kit includes SuperScript IV Reverse Transcriptase (RT), a proprietary MMLV mutant that produces robust and reliable cDNA synthesis in RT reactions, and ezDNase to remove potential genomic DNA contamination. The SuperScript IV synthesis system is significantly improved over the SuperScript III-based system in inhibitor resistance, processivity, and reaction speed, while retaining all the benefits of the previous system, including increased thermostability, highly efficient full-length cDNA synthesis, and reduced RNase activity. The SuperScript IV system is designed to provide reliable, consistent, and fast cDNA synthesis in the presence of inhibitors found in a wide variety of samples that cause other currently available RTs to perform inefficiently. This system is also available without ezDNase.

The SuperScript IV First-Strand Synthesis System contains all components needed for RT reactions, plus an additional control gene and primers, and provides the flexibility to customize the RT set-up to fit the needs of your application. For even better performance, ezDNase for genomic DNA removal is included. The extremely simplified genomic DNA removal step dramatically reduces the time of the entire reverse transcription protocol and reduces possible variation in gene expression due to RNA loss or damage during conventional DNase treatment. The SuperScript IV synthesis system with ezDNase is the top choice for performance and flexibility for RT-PCR applications. We recommend it for all RT-PCR applications, especially when reproducibility and reliability is the primary concern and when inhibitors in the RNA sample may interfere with cDNA synthesis, leading to biases in gene expression studies.

Features of the SuperScript IV First-Strand Synthesis System include:
• Significantly improved resistance to a variety of inhibitors that can interfere with cDNA synthesis
• Robust and specific cDNA synthesis in a wide range of sample types
• A faster reverse transcriptase reaction that reduces incubation time from >50 min to 15 min including a simplified genomic DNA removal step
• Significantly better processivity compared to SuperScript III RT

Enzyme source
Purified from E. coli expressing the pol gene of M-MLV, modified to increase thermostability and half-life, processivity, inhibitor resistance and to reduce RNase H activity.

Performance and quality testing
Endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product.

Unit definition
One unit of SuperScript IV RT is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min at 37°C using poly(A) oligo(dT)12-18 as a template/primer.

Unit reaction conditions
50 mM Tris-HCl (pH 8.3), 4 mM MgCl2, 10 mM DTT, 50 mM KCl, 0.5 mM dTTP, 0.4 MBq/mL [3H]-dTTP, 0.4 mM poly(A) oligo (dT)12-18 and enzyme in 20 μL for 10 min at 37°C.

SuperScript™ IV One-Step RT-PCR System with ezDNase™ (Invitrogen™)

The Invitrogen SuperScript IV One-Step RT-PCR System with ezDNase combines the high processivity of SuperScript IV Reverse Transcriptase (RT) with the high-fidelity of Platinum SuperFi DNA Polymerase to provide unmatched one-step RT-PCR performance. With this convenient one-step formulation and gene-specific primers, you can perform both cDNA synthesis from total RNA or mRNA and PCR amplification in a single tube.

Features of the SuperScript IV One-Step RT-PCR System with ezDNase include:
• Superior performance with sensitivity down to 0.01 pg RNA, target length up to 13.8 kb, and the fastest one-step RT-PCR protocol
• Reliable target detection even with suboptimally pure RNA samples
• Two-phase hot-start activation mechanism for high sensitivity, specificity, and room temperature setup
• Fast and easy gDNA removal for superior accuracy and confidence in results

SuperScript IV RT is a proprietary MMLV RT mutant with increased processivity, thermostability, and inhibitor resistance, as well as reduced RNase H activity, for sensitive and efficient full-length cDNA synthesis. Platinum SuperFi DNA Polymerase is a proofreading DNA polymerase that combines superior fidelity with trusted Platinum hot-start technology for the highest success in PCR. The combination of these two engineered enzymes produces enhanced RT-PCR yields and fidelity, as well as amplification of longer templates compared to other one-step RT-PCR kits. Inclusion of ezDNase enzyme for use upstream of the RT step further accelerates the RT-PCR workflow through an extremely simplified genomic DNA removal step. (This one-step RT-PCR system is also available without ezDNase.)

SuperScript™ One-Step RT-PCR System with Platinum™ Taq DNA Polymerase (Invitrogen™)

The SuperScript One-Step RT-PCR System with Platinum Taq DNA Polymerase is designed for the convenient end-point detection and analysis of RNA molecules by RT-PCR. This system combines RT/ Platinum Taq Mix and 2X Reaction Mix. The RT/ Platinum Taq Mix contains a mixture of SuperScript II Reverse Transcriptase and Platinum Taq DNA Polymerase. The 2X Reaction Mix consists of a proprietary buffer system optimized for reverse transcription and PCR amplification, Mg2+ optimized for universal use, deoxyribonucleotide triphosphates, and stabilizers. All components for cDNA synthesis and PCR are combined in a single tube with gene-specific primers and target RNA. Reverse transcription automatically follows PCR cycling without additional steps.

Note: For superior one-step RT-PCR performance, we recommend the SuperScript IV One-Step RT-PCR System or SuperScript IV One-Step RT-PCR System with ezDNase. The SuperScript IV One-Step RT-PCR System combines the high processivity of SuperScript IV Reverse Transcriptase with the high-fidelity of Platinum SuperFi DNA Polymerase to provide unmatched product yields, specificity, and sensitivity in less time and for a broad target range, even with suboptimally pure RNA samples.

Verso 1-Step RT-PCR Kit ReddyMix, with ThermoPrime Taq (Thermo Scientific™)

Thermo Scientific Verso 1-Step RT-PCR Kit ReddyMix (with ThermoPrime Taq) is a complete set of RT-PCR reagents that combines everything needed for RT-PCR. The Verso RT enzyme delivers high processivity and can be used at temperatures up to 57°C, delivering effective transcription through difficult RNA secondary structures. Verso also shows a better dynamic range than standard AMV or M-MuLV enzymes, with increases in template concentration giving rise to corresponding increases in product generated over a wider range of starting material concentrations.

An optional RT Enhancer is provided with the kit and can be used during the RT step to remove any DNA contamination, eliminating the need for DNase I treatment.

This Verso 1-step RT-PCR system comes with the ReddyMix option, where the master mix contains a dye and precipitant that allow RT-PCR reactions to be loaded directly onto an electrophoresis gel. The red dye migrates between bromophenol blue and xylene cyanol at approximately 300 bp, depending on agarose concentration.

Highlights:

• ReddyMix formulation—eliminates post-PCR steps required for electrophoresis
• Eliminate the need for DNase treatments—RT Enhancer (optional) prevents gDNA carry-over
• RT and PCR steps performed in single sealed tube—convenience and reduced contamination risk
• Verso enzyme is highly sensitive in a broad dynamic range—accurately detect RNA input from 1 pg to 1 µg
• Wide working temperature range (42 to 57°C) for success with GC-rich and other difficult templates
• RNase Inhibitor included in RT enzyme mix

Applications:

• Gene expression analysis
• Viral/bacterial detection
• 1-step RT-PCR
• High throughput applications

Related Products
Verso 1-Step RT-PCR Kit ReddyMix, with ThermoPrime Taq
Verso 1-Step RT-PCR Kit, with Thermo-Start Taq (Hot Start)