RevertAid H Minus Reverse Transcriptase (200 U/μL)
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RevertAid H Minus Reverse Transcriptase (200 U/μL)
Thermo Scientific™

RevertAid H Minus Reverse Transcriptase (200 U/μL)

Thermo Scientific RevertAid H Minus Reverse Transcriptase is a recombinant M-MuLV RT. The enzyme possesses an RNA-dependent and DNA-dependent polymeraseRead more
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Catalog NumberQuantity
EP045110,000 Units
EP04525 x 10,000 Units
Catalog number EP0451
Price (USD)
165.00
Each
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Quantity:
10,000 Units
Request bulk or custom format
Price (USD)
165.00
Each
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Thermo Scientific RevertAid H Minus Reverse Transcriptase is a recombinant M-MuLV RT. The enzyme possesses an RNA-dependent and DNA-dependent polymerase activity, but lacks RNase H activity due to a point mutation in the RNase H domain. RevertAid H Minus Reverse Transcriptase does not degrade RNA in RNA-DNA hybrids during synthesis of the first strand cDNA, and therefore high yields of full-length cDNA from long templates are obtained. RevertAid H Minus Reverse Transcriptase maintains activity over a wide temperature range (42–50°C).

Features of RevertAid H Minus Reverse Transcriptase include:
• High yields of full-length first strand cDNA up to 13 kb
• Optimum activity at 42–45°C
• Active up to 55°C
• Incorporates modified nucleotides (e.g., Cy3-, Cy5-, rhodamine-, aminoallyl-, and fluorescein-labeled nucleotides)

Applications
• First strand cDNA synthesis for RT-PCR and real-time RT-PCR
• Reverse transcription at elevated temperatures to reduce effects of secondary structure
• Synthesis of cDNA for cloning and expression
• Generation of labeled cDNA probes for microarrays
• DNA labeling
• Analysis of RNA by primer extension

For Research Use Only. Not for use in diagnostic procedures.
Specifications
FormatStand-alone Enzyme
Reaction Speed60 min.
TechniqueReverse Transcription
Optimal Reaction Temperature42°C to 45°C
Quantity10,000 Units
Reverse TranscriptaseRevertAid H Minus
Ribonuclease H ActivityReduced
Shipping ConditionDry Ice
Concentration200 U/μL
Final Product TypeFirst-Strand cDNA
Reaction FormatSeparate components
Reagent TypeReverse Transcription
Size (Final Product)Up to 13 kb
Starting MaterialRNA
Unit SizeEach
Contents & Storage

• RevertAid H Minus Reverse Transcriptase (200 U/μL)
• 5X Reaction Buffer

Store at –20°C.

Frequently asked questions (FAQs)

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. It is also recommended to use RNase H-minus RTs for template-independent addition of C nucleotides. In contrast, reverse transcriptases with intrinsic RNase H activity are often favored in qPCR applications.

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 possess intrinsic TdT activity although at varying degrees depending upon the reaction conditions. For addition of template-independent C nucleotides (as for SMART and RACE experiments), this specific TdT activity can be induced by Mn2+. We would recommend Maxima H- or RevertAid H- minus RTs for this purpose.

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. It is also recommended to use RNase H Minus RTs for template-independent addition of C nucleotides. In contrast, reverse transcriptases with intrinsic RNase H activity are often favored in qPCR applications.

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