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  7. Thermo Scientific™ Maxima H Minus Double-Stranded cDNA Synthesis Kit

Thermo Scientific™ Maxima H Minus Double-Stranded cDNA Synthesis Kit

Catalog No. FERK2561
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FERK2561 10 Reactions
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Catalog No. FERK2561 Supplier Thermo Scientific™ Supplier No. K2561
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Description

Includes

First Strand Enzyme Mix, 4X First Strand Reaction Mix, Second Strand Enzyme Mix, 5X Second Strand Reaction Mix, 0.5M EDTA ( pH 8.0), RNase I (10 u/μL), Control RNA (0.5μg/μL), Oligo(dT)18 Primer, 100μM, 0.5μg/μL, Random Hexamer Primer, 100μM, 0.2μg/μL,Water, nuclease-free, Detailed Protocol

The Thermo Scientific Maxima H Minus Double-Stranded cDNA Synthesis Kit is a complete system for efficient synthesis of double-stranded cDNA from total RNA or mRNA.

Thermo Scientific Maxima H Minus Double-Stranded cDNA Synthesis Kit is a complete system for efficient synthesis of double-stranded cDNA from total RNA or mRNA. First- and second-strand cDNA synthesis reactions are performed in the same tube without the need for intermediate organic extraction or ethanol precipitation steps. This convenient single-tube format speeds up the synthesis procedure and maximizes cDNA recovery. The kit contains premixed components to reduce the number of pipetting steps necessary to complete the procedure.

First- and second-strand cDNA synthesis reactions are performed in the same tube without the need for intermediate organic extraction or ethanol precipitation steps. This convenient single-tube format speeds up the synthesis procedure and maximizes cDNA recovery. The kit contains pre-mixed components to reduce the number of pipetting steps necessary to complete the procedure.

Features of the Maxima H Minus Double-Stranded cDNA Synthesis Kit include:

  • Efficient synthesis of full-length double-stranded cDNA
  • Fast—procedure completed in less than two hours
  • Convenient—pre-mixed components
  • Complete—includes all primers, controls and residual RNA removal reagents

Applications

  • Full-length double-stranded blunt-end cDNA synthesis for cloning
  • Double-stranded cDNA library construction

Order Info

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

Specifications

Format 10 reactions
Optimal Reaction Temperature 50°-55° C
Product Line Maxima
Product Type cDNA Synthesis Kit
Quantity 10 Reactions
Reverse Transcriptase Maxima H Minus
For Use With (Application) Cloning
Final Product Type Double Stranded cDNA
No. of Reactions 10 Reactions
Reaction Format Separate Components
Sample Type RNA
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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. We also recommended using RNase H-minus RTs for template-independent addition of C nucleotides.

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. 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.