Maxima™ H Minus cDNA Synthesis Master Mix
Maxima™ H Minus cDNA Synthesis Master Mix
Thermo Scientific™

Maxima™ H Minus cDNA Synthesis Master Mix

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 NumberIncludesNo. of Reactions
M1661Kit only50 Reactions
M1662Kit only200 Reactions
M1681Kit with dsDNase50 Reactions
M1682Kit with dsDNase200 Reactions
Catalog number M1661
Price (USD)
363.65
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450.00
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Includes:
Kit only
No. of Reactions:
50 Reactions
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Price (USD)
363.65
Online Exclusive
450.00
Save 86.35 (19%)
Each
Add to cart
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 convienient 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)

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
Concentration5X
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
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. 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.

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.

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