missing translation for 'onlineSavingsMsg'
Learn More
  1. Home
  2. Products
  3. PCR Equipment and Supplies
  4. PCR Supplies
  5. Reverse Transcription Products
  6. cDNA Synthesis Kits and Master Mixes
  7. Thermo Scientific™ Maxima™ H Minus cDNA Synthesis Master Mix

Thermo Scientific™ Maxima™ H Minus cDNA Synthesis Master Mix

Catalog No. FERM1682
Change view
Click to view available options
Includes:
Kit only
Kit with dsDNase
No. of Reactions:
200 Reactions
50 Reactions
4 product options available for selection
Product selection table with 4 available options. Use arrow keys to navigate and Enter or Space to select.
Catalog No. Includes No. of Reactions
FERM1682 Kit with dsDNase 200 Reactions
FERM1661 Kit only 50 Reactions
FERM1662 Kit only 200 Reactions
FERM1681 Kit with dsDNase 50 Reactions
Use arrow keys to navigate between rows. Press Enter or Space to select a product option. 4 options available.
4 options
Catalog No. FERM1682 Supplier Thermo Scientific™ Supplier No. M1682
Only null left

Description

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.

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)

Specifications

Content And Storage

• Maxima H Minus cDNA Synthesis Master Mix (1 x 800 μL)
• Maxima H Minus cDNA Synthesis Master Mix 'No RT' Control (1 x 200 μL)
• dsDNase (1 x 200 μL)
• 10X dsDNase buffer (1 x 400 μL)
• Nuclease-free water (1 x 1.25 mL)

Store at –5 to –30°C.
Format Master Mix
GC-Rich PCR Performance High
Reaction Speed 30 min.
Technique Reverse Transcription
Optimal Reaction Temperature 50°C
Quantity Each
Reverse Transcriptase Maxima H Minus
Ribonuclease H Activity Reduced
Shipping Condition Dry Ice
For Use With (Application) Real Time PCR (qPCR), RT-PCR
Final Product Type First-Strand cDNA
No. of Reactions 200 Reactions
Reaction Format Premixed Components
Reagent Type Reverse Transcription
Size (Final Product) Up to 20 kb
Starting Material RNA
Includes Kit with dsDNase
Show More Show Less

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.
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.