Thermo Scientific™ RevertAid RT Reverse Transcription Kit

Product Code.	Quantity	unitSize
15255146	500 reactions	500 reactions

Product Code. 15255146 Supplier Thermo Scientific™ Supplier No. K1691

Description

The Thermo Scientific RevertAid RT Kit is a complete system for efficient synthesis of first strand cDNA from mRNA or total RNA templates.

The Thermo Scientific RevertAid RT Kit is a complete system for efficient synthesis of first strand cDNA from mRNA or total RNA templates. The kit uses RevertAid Reverse Transcriptase (RT), which has lower RNase H activity compared to AMV reverse transcriptase. The enzyme maintains activity at 42–50°C and is suitable for synthesis of cDNA up to 13 kb. The recombinant RiboLock RNase Inhibitor, supplied with the kit, effectively protects RNA from degradation at temperatures up to 55°C.

First strand cDNA synthesized with this system can be directly used as a template in PCR or real-time PCR. It is also ideal as a first step for second strand cDNA synthesis or linear RNA amplification. Radioactively and non-radioactively labeled nucleotides can be incorporated into first strand cDNA for use as a probe in hybridization experiments, including microarrays.

Features of the RevertAid RT Kit include:
• Full-length first strand cDNA up to 13 kb
• Optimum reaction temperature 42°C
• Complete kit includes all the components for RT reactions

Applications
• First strand cDNA synthesis for RT-PCR and RT-qPCR
• Construction of full length cDNA libraries
• Antisense RNA synthesis

Order Info

Orders placed Monday through Wednesday ship next day; Thursday and Friday orders ship the following Monday.

Specifications

• Content And Storage:

  • RevertAid Reverse Transcriptase
  • Ribolock RNase Inhibitor
  • 5X Reaction Buffer
  • dNTP Mix
  • Random Hexamer Primer
  • Nuclease-free water
  
  Store at –20°C.
  

  
  
• Format: Kit
• Reaction Speed: 60 min.
• Technique: Reverse Transcription
• Optimal Reaction Temperature: 42°C
• Quantity: 500 reactions
• Reverse Transcriptase: RevertAid
• Ribonuclease H Activity: Yes
• Shipping Condition: Dry Ice
• For Use With (Application): Real Time PCR (qPCR)
• Final Product Type: First-Strand cDNA
• No. of Reactions: 500 Reactions
• Reaction Format: Separate components
• Reagent Type: Reverse Transcription
• Size (Final Product): Up to 13 kb
• Starting Material: RNA

Frequently Asked Questions (FAQs)

What steps should I take while performing first strand cDNA synthesis using low purity template (e.g., inhibitors in RNA sample)?

Trace amounts of reagents used in RNA purification protocols may remain in solution and inhibit first-strand synthesis, e.g., SDS, EDTA, guanidine salts, phosphate, pyrophosphate, polyamines, spermidine. To remove trace contaminants, we recommend re-precipitating the RNA with ethanol and washing the pellet with 75% ethanol, or re-purifying the RNA.

For reverse transcription, how important is the quality of RNA template?

RNA purity and integrity are essential for synthesis and quantification of cDNA. Always assess the integrity of RNA prior to cDNA synthesis. Use freshly prepared RNA. Multiple freeze/thaw cycles of the RNA sample and synthesized cDNA is not recommended. Avoid RNase contamination and discard low quality RNA.

When should I choose regular RevertAid RT or Maxima RT vs. RevertAid H Minus RT or Maxima H Minus RT?

It is generally beneficial to minimize RNase H activity when aiming to produce long transcripts for cDNA cloning. RNase H degrades RNA from RNA-DNA duplexes, which can result in truncated cDNA during reverse transcription of long mRNA. We also recommended using RNase H Minus RTs for template-independent addition of C nucleotides. In contrast, reverse transcriptases with intrinsic RNase H activity are often favored in 1-step RT-qPCR applications.

What is the fidelity of RevertAid and Maxima reverse transcriptases?

The fidelity of RevertAid and Maxima reverse transcriptases is the same as that of wild-type M-MuLV RT, which is in the range of 1 error per 15,000-27,000 nucleotides synthesized.

Do Thermo Scientific reverse transcriptases (RevertAid RT, RevertAid H Minus RT, Maxima RT, and Maxima H Minus RT) possess terminal deoxynucleotidyl (TdT) activity?

All Thermo Scientific reverse transcriptases (RevertAid RT, RevertAid H Minus RT, Maxima RT, and Maxima H Minus RT) possess intrinsic TdT activity, although at varying degrees depending upon the reaction conditions. We recommend specialized SuperScript IV Template Switching RT Master Mix for high efficiency in applications requiring template switching RT.

For Research Use Only. Not for use in diagnostic procedures.