Reverse Transcriptases Overview
A major factor to consider in reverse transcription polymerase chain reaction (RT-PCR) is the choice of the appropriate reverse transcriptase.Click on the application below to find the recommended product:
> One-Step RT-PCR on Conventional Thermal Cyclers
> One-Step RT-PCR on Real-Time Instruments
> Two-Step RT-PCR on Conventional Thermal Cyclers
> Two-Step RT-PCR on Real-Time Instruments
Additional Reverse Transcriptases
In addition to the above recommended products for one- and two-step RT-PCR, Roche has developed and optimized a wide variety of conventional enzymes and enzyme blends. Each of these products has different enzymatic properties and one may be more suitable for a specific experiment than the other. The information given below outlines the individual properties of each enzyme and may help you find the optimum enzyme or enzyme blend for your specific application.
One-step RT-PCR - offers faster reactions, reduced risk of contamination, and improved sensitivity
Tth DNA Polymerase | Titan One Tube RT-PCR System | Titan One Tube RT-PCR Kit | |
Composition | Thermostable DNA polymerase with intrinsic reverse transcriptase activity. | AMV reverse transcriptase and Expand High Fidelity Enzyme Blend. | |
Product Size | Up to 1 kb | Up to 6 kb | |
Priming | Specific | Specific | |
Reaction Temperature | +55 to +70°C | +45 to +60°C | |
RNase H Activity | - | ++ | |
Ion Requirement | Mn2+ | Mg2+ | |
Sensitivity - Total RNA | + | 1 pg to 1 µg | |
Sensitivity - mRNA | + | ++ | |
Sensitivity - Viral RNA | + | + | |
Sensitivity - RNA Rich in Secondary Structures | + | + | |
Specificity | + | + | |
Fidelity | Highly processive 5´-3´ polymerase, lacking 3´-5´ exonuclease or proofreading activity. | + | |
Full-Length cDNA | - | ++ | |
Incorporation of Modified Nucleotides | Yes | Yes | |
Carryover Prevention | Yes | No | |
Buffer System | Both a PCR and RT-PCR buffer are included. | A single reaction buffer is included. | |
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Two-step RT-PCR - offers maximum flexibility with respect to choice of primers, enzyme, temperature, and analysis of multiple transcripts
Reverse Transcriptase M-MuLV | Expand Reverse Transcriptase | Reverse Transcriptase AMV | First Strand cDNA Synthesis Kit | cDNA Synthesis System | |
Product Size | 10 kb | 14 kb | 12 kb | ||
Priming | Specific Oligo(dT) Random hexamer | Specific Oligo(dT) Random hexamer | Specific Oligo(dT) Random hexamer | ||
Reaction Temperature | +37°C | +42°C (up to +50°C) | +42°C (up to +60°C) | ||
RNase H Activity | + | - | ++ | ||
Sensitivity - Total RNA | + | ++ | + | ||
Sensitivity - mRNA | + | ++ | ++ | ||
Sensitivity - Viral RNA | + | + | + | ||
Sensitivity - RNA Rich in Secondary Structures | +(+) | + | + | ||
Specificity | Not determined | + | + | ||
Fidelity | + | + | + | ||
Full-Length cDNA | + | +++ | ++ | ||
Incorporation of Modified Nucleotides | Yes | Yes | Yes | ||
Carryover Prevention | Not determined | Not determined | Not determined | ||
Buffer System | A single reaction buffer is included. Separate vials of DMSO, DTT, and Betain for optimization of reaction conditions are also included. | A single reaction buffer is included. A separate vial of DTT for optimization of reaction conditions is also included. | All necessary reagents are included. | ||
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