Thermo Scientific™

Maxima™ H Minus cDNA Synthesis Master Mix

Name: Maxima™ H Minus cDNA Synthesis Master Mix
SKU: M1661

Thermo Scientific Maxima H Minus cDNA Synthesis Master Mix, available with or without dsDNase, provides all cDNA synthesis reaction components in a convenient one-tube master mix.

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Catalog Number	Includes	No. of Reactions
M1661	Kit only	50 Reactions
M1662	Kit only	200 Reactions
M1681	Kit with dsDNase	50 Reactions
M1682	Kit with dsDNase	200 Reactions

4 Options

Catalog number M1661

Price (MXN)

Includes:

Kit only

No. of Reactions:

50 Reactions

Request bulk or custom format

Thermo Scientific Maxima H Minus cDNA Synthesis Master Mix, available with or without dsDNase, provides all cDNA synthesis reaction components in a convenient one-tube master mix. It is optimized for highly efficient cDNA synthesis for two-step quantitative RT-PCR (RT-qPCR) applications and reproducible cDNA synthesis at elevated temperatures (50–65°C) within 30 minutes.

The master mix contains Maxima H Minus Reverse Transcriptase (RT) and RiboLock RNase Inhibitor. Maxima H Minus RT is an advanced enzyme derived from M-MuLV RT by in vitro evolution. The enzyme features high thermostability, robustness, processivity, increased cDNA synthesis rate, and lack of RNase H activity. The recombinant RiboLock RNase Inhibitor effectively protects RNA template from degradation by RNases A, B, and C at temperatures up to 55°C. The master mix also contains reaction buffer, dNTPs, oligo (dT)₁₈, and random hexamer primers. A separate tube of No-RT control mix is also provided for convenient RT negative control.

Both kits are capable of reproducible cDNA synthesis at elevated temperatures (50°C–65°C). The synthesis reaction is typically complete in 15–30 minutes. The included double-strand specific DNase in the Maxima H Minus cDNA Synthesis Master Mix with dsDNase allows removal of genomic DNA from RNA samples in two minutes without affecting the quality or quantity of RNA. This dsDNase is also available separately (Cat. No. EN0771).

Features of Maxima H Minus cDNA Synthesis Master Mix include:

One-tube master mix helps reduce pipetting steps and enhanced consistency in RT-qPCR results
Increased RT efficiency across a wide range of input RNA amounts and gene targets
High thermostability to allow RT reactions at 50–65°C temperature range
Integrated step of genomic DNA removal from RNA samples (Maxima H Minus cDNA Synthesis Master Mix with dsDNase only)

Full-length cDNA synthesis for long transcripts and ORFs
Reverse transcription at elevated temperatures to reduce effects of secondary structure
cDNA cloning and construct generation
RNA-seq & single cell RNA-seq library prep
Isoform and transcript characterization
Template-switching-based full-length RNA-seq

Additional information about reaction components

The No RT control mix contains all the components in the Maxima H Minus cDNA Synthesis Master Mix except RT. The presence of a PCR product in the No RT control reaction indicates that the reaction is contaminated with DNA. To further enhance genomic DNA elimination efficiency, template RNA incubation with dsDNase (Cat. No. EN0771) is strongly recommended.
Nuclease-free water is provided for reaction setup and dilution of sample DNA. The absence of exodeoxyribonucleases, ribonucleases, and phosphatases has been confirmed by appropriate quality tests.
To further enhance genomic DNA elimination efficiency, template RNA incubation with the included dsDNase is strongly recommended. (Maxima H Minus cDNA Synthesis Master Mix with dsDNase only)
The dsDNase is used for rapid and safe removal of contaminating genomic DNA from RNA samples. dsDNase is easily inactivated by moderate heat treatment (55°C). It allows for a dramatically simplified workflow that combines genomic DNA elimination and cDNA synthesis into one-tube procedure. (Maxima H Minus cDNA Synthesis Master Mix with dsDNase only)

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

Specifications

Final Product TypeFirst-Strand cDNA

FormatMaster Mix

IncludesKit only

No. of Reactions50 Reactions

Optimal Reaction Temperature50°C

QuantityEach

Reaction FormatPremixed Components

Reagent TypeReverse Transcription

Reverse TranscriptaseMaxima H Minus

Ribonuclease H ActivityReduced

Shipping ConditionDry Ice

Size (Final Product)Up to 20 kb

Starting MaterialRNA

TechniqueReverse Transcription

Concentration5X

For Use With (Application)Real Time PCR (qPCR), RT-PCR

GC-Rich PCR PerformanceHigh

Reaction Speed30 min.

Unit SizeEach

Contents & Storage

• Maxima H Minus cDNA Synthesis Master Mix (200 μL)
• Maxima H Minus cDNA Synthesis Master Mix 'No RT' Control (200 μL)
• Nuclease-free water (1.25 mL)

Store at –5 to –30°C.

Frequently asked questions (FAQs)

Why is there no heat inactivation step for the dsDNase in the protocol for Maxima H Minus cDNA Synthesis Master Mix (Cat. No. M1681)?

The novel dsDNase enzyme degrades double-stranded genomic DNA. However, it has no effect on the single-stranded cDNA or the RNA-DNA hybrid. Therefore, the enzyme mix for reverse transcription can be added directly to the dsDNase treated RNA.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

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.

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.