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Ordering

Cat. No.
Size
BIO-65111
10,000 Units
BIO-65112
4 x 10,000 Units

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Description

Thermostable Reverse Transcriptase is a robust thermostable MMLV-RT, designed to reduce RNase H activity and increase thermal stability. The enzyme offers higher cDNA yields and improved efficiency on GC-rich target RNAs and RNA with high secondary structure, such as viral RNA sequences.

Product Highlights

  • Stable - Reverse transcriptase activity up to 60°C
  • Robust - Ideal for RNA with high secondary structure such as viral genomes
  • Sensitive - Detection of low copy number RNA targets
  • Glycerol-free - Formulation compatible with lyophilization and air-drying applications
  • Fast - Ideal for developing fast, highly reproducible RT-qPCR assays

Product Description

Traditional Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV-RT) is not thermostable and can only maintain its enzymatic activity at relatively low temperatures (up to 50°C). However for cDNA synthesis, a higher reaction temperature is desirable as it reduces RNA secondary structures which can inhibit reverse transcription and it minimizes nonspecific primer binding.

Meridian Thermostable Reverse Transcriptase has higher thermal stability and reduced RNase H activity. The enzyme can be used to synthesize first-strand cDNA at temperatures up to 60°C which improves the cDNA yield from difficult RNA targets that require higher temperature to denature strong secondary structures.

Main

Specification

Components

Reagent

10,000 units

4 x 10,000 units

Thermostable Reverse Transcriptase

50 µL

4 x 50 µL

5x Reaction Buffer

1 mL

4 x 1 mL

Storage & Stability

Thermostable Reverse Transcriptase can be stored for up to 12 months at -20°C. Repeated freeze/thaw cycles should be avoided.

When stored under the recommended conditions and handled correctly, full activity of the kit is retained until the expiry date printed on the outer box label.

Shipping conditions

On dry or blue ice



FAQs

For SensiFAST cDNA Synthesis Kit: The TransAmp Buffer already contains reverse transcriptase enhancers that reduce complex RNA secondary structure. For templates that have a high degree of structure, such as viral RNA and some plant RNA, we suggest using an additional 15 minute 48°C incubation step as mentioned in the manual. For Tetro cDNA Synthesis Kit: For samples with high secondary structure we suggest to denature the RNA template with primers prior to the reaction set-up. Furthermore, it is necessary to optimize the incubation time and temperature. The temperature of the RT step may be raised up to a maximum of 48°C.

The necessary amount of cDNA template for PCR or qPCR reactions may vary from the range of pg up to several hundred ng. It depends among other things on the analyzed target, the used application and sample characteristics. If you do not have empirical values for your assays please use the recommended amounts in the manuals of your PCR or qPCR assays and optimize if necessary.

It is not possible for the SensiFAST cDNA Synthesis Kit. Random Hexamer and oligo dT primer are already included in the optimized TransAmp buffer. It is possible to use gene specific primer for the Tetro cDNA Synthesis Kit to generate cDNA for specific target.

The SensiFAST cDNA Synthesis Kit has been designed for rapid generation and unbiased coverage of cDNA, especially for subsequent quantification with real-time PCR. The optimized 5x TransAmp buffer includes all necessary components for cDNA synthesis, including a unique blend of oligo dT and random hexamer primers and the RiboSafe RNase Inhibitor. The Tetro cDNA Synthesis Kit is the more flexible solution. You selects the desired setup, especially the desired RT primer. The Tetro Kit is optimized to work over a wide range of RNA concentrations and to obtain full length cDNA, which can be used for two-step PCR assays as well. Tetro is the recommended Kit for the generation of cDNA libraries and cloning.