Thermo Scientific Maxima Reverse Transcriptase and Thermo Scientific Maxima H Minus Reverse Transcriptase were engineered to offer high thermostability, processivity, and sensitivity, enabling consistent and reliable results in RT-PCR, RT-qPCR, and RNA sequencing applications. Both reverse transcriptase (RT) enzymes exhibit strong tolerance to common inhibitors, helping ensure robust performance across challenging samples.

Both Maxima RTs were developed through in vitro evolution of M-MuLV RT. While Maxima RT possesses RNase H activity, Maxima H Minus RT is RNase H–deficient, which prevents RNA degradation during reverse transcription, allowing synthesis of longer cDNA and higher yields. These RTs are available in a variety of formats including stand-alone enzymes, kits, and master mixes.


Features of Maxima Reverse Transcriptases

  • High yields of cDNA over a wide temperature range (up to 65°C): Improve cDNA synthesis from challenging RNA samples, including GC-rich or highly structured transcripts, for greater confidence in downstream analysis.
  • Strong linearity and reproducibility in two-step RT-qPCR: Achieve consistent, quantitative gene expression data across experiments, improving confidence in experimental outcomes.
  • Long cDNA transcripts—up to 20 kb: Generate full-length cDNA from long RNA targets for comprehensive gene expression and transcript characterization studies. Reduced Rnase H activity of Maxima H Minus RT is suitable for efficient RT-PCR with long amplification targets.
  • Sensitivity—as low as 1 pg of total RNA: Obtain meaningful results from limited samples, enabling analysis of rare, low-input, or low-purity RNA samples.
  • Improved performance in the presence of inhibitors that can interfere with cDNA synthesis: Maintain reliable performance across diverse sample types and purification methods, reducing the risk of failed reactions and repeat experiments.
  • Accurate results with no false signal from gDNA contamination—integrated dsDNase and no-RT control: Increase confidence in your data by removing genomic DNA interference, helping ensure that detected signals reflect true RNA expression rather than contamination.

Maxima Reverse Transcriptases Applications

  • Full-length cDNA synthesis for long transcripts and ORFs
  • RT-qPCR for gene expression analysis and high-throughput screening
  • cDNA cloning and construct generation
  • RNA-seq and single cell RNA-seq library prep
  • Isoform and transcript characterization
  • SMART-seq, RACE, and other template switching applications

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Product offerings

Maxima RTs and Maxima H Minus RTs are available in a variety of formats including stand-alone enzymes, kits, and master mixes.

  • Standalone enzymes for maximum flexibility in reaction setup.
  • Complete kits/master mixes for efficient and reproducible first-strand cDNA synthesis. These kits are also available in formats with dsDNase. The dsDNase step is completed in two minutes and allows for an easy gDNA removal prior to reverse transcription.
ProductFormatCatalog number 
Maxima Reverse TranscriptaseStand-alone enzymeEP0741Order now
First Strand cDNA Synthesis KitK1641
(Kit only)		Order now
K1671
(Kit with dsDNase)		Order now
Maxima H Minus Reverse TranscriptaseStand-alone enzymeEP0751Order now
First Strand cDNA Synthesis KitK1651
(Kit only)		Order now
cDNA Synthesis Master MixM1681
(Kit with dsDNase)		Order now

Maxima H Minus Reverse Transcriptase: Performance data

For routine RT-qPCR applications, Maxima H Minus Reverse Transcriptase enables efficient cDNA synthesis over a wide range of input RNA amounts, helping provide sensitive and accurate quantification of cDNA. In addition, for advanced RNA workflows such as RNA-sequencing and RT-qPCR of long targets, Maxima H Minus Reverse Transcriptase enables efficient full-length cDNA synthesis even from challenging samples.

Compared to tested reverse transcriptases, Maxima H minus RT produced higher yields of full-length cDNA over a wide temperature range (Figure 1). Its tolerance to high reaction temperatures allows efficient transcription of RNA regions with extensive secondary structures and helps to improve primer specificity, thereby increasing overall yields.

Agarose gel electrophoresis results comparing cDNA synthesis performance of seven reverse transcription kits.

Figure 1. Reliable performance across temperature range and efficient cDNA synthesis. cDNA was synthesized from Invitrogen Millenium™ RNA Markers by incubating samples at different temperatures for 10 min, using (A) Maxima H Minus Reverse Transcriptase and reverse transcriptases from other suppliers: (B) NEB LunaScript® RT SuperMix Kit, (C) NEB ProtoScript® II First Strand cDNA Synthesis Kit, (D) BioRad Reliance Select cDNA Synthesis Kit, (E) BioRad iScript™ Advanced cDNA Synthesis Kit, (F) Quanta qScript Ultra SuperMix and (G) Quanta qScript Ultra Flex Kit. cDNA products were detected by agarose gel electrophoresis.

Maxima H Minus Reverse Transcriptase efficiently synthesizes cDNA from a wide range of input RNA, has higher yields, and better linearity in cDNA synthesis compared to commercially available reverse transcriptases, making it a good choice for RT-qPCR experiments (Figure 2, 3 and 4).

Figure 2. High sensitivity and yield. Maxima H Minus Reverse Transcriptase compared with other commercially available RTs using 5 orders of magnitude (10 pg–1 μg) of total HeLa RNA input in RT-qPCR. qPCR performance was evaluated with Applied Biosystems TaqMan Fast Advanced Master Mix. The results demonstrate strong linearity, with high correlation coefficient (R² = 0.996) and robust amplification efficiency (eff% = 90%). Compared to other commercial reverse transcriptases, Maxima H Minus RT delivers earlier Ct values (red line), indicating higher cDNA yield and detection sensitivity.

Figure 3. Maxima H Minus RT delivers earlier Ct values compared to other commercial RTs. Reverse transcription efficiency of Maxima H Minus RT was compared with commercial RTs by RT-qPCR using 100 ng of total HeLa RNA as input. qPCR performance was evaluated with Applied Biosystems TaqMan Fast Advanced Master Mix and results are shown as ΔCt [ΔCt = Ct competitor − Ct Maxima H Minus RT]. Bars represent the Ct delay observed with competitor kits relative to Maxima H Minus Reverse Transcriptase across multiple gene targets. Lower Ct values obtained with Maxima H Minus indicate efficient cDNA synthesis and consistent sensitivity across the genes tested.

 

four RT-qPCR amplification plots comparing the performance of Maxima H Minus cDNA Synthesis Master Mix with several commercial reverse transcription kits

Figure 4. Enhanced transcription efficiency of Maxima H Minus cDNA Synthesis Master Mix. The Maxima H Minus cDNA Synthesis Master Mix demonstrates better efficiency over a wide range of input RNA amounts when compared to competitor’s RTs. Amplification of the human 18S RNA gene was performed on 10-fold serial dilutions of HeLa total RNA (1 μg to 0.1 pg). First-strand cDNA was generated using the Maxima H Minus cDNA Synthesis Master Mix, Thermo Scientific Maxima First Strand cDNA Synthesis Kit for RT-qPCR, and reverse transcriptases from four other suppliers. cDNA was amplified using the Luminaris Probe qPCR Master Mix, low ROX on the ViiA 7 Real-Time PCR System. Amplification plots indicate variation of ΔRn with cycle number.

Maxima H Minus Reverse Transcriptase demonstrates up to 50x higher processivity than wild-type M-MuLV RT, enabling efficient synthesis of full-length cDNA from a wide range of RNA templates, including very long transcripts up to 20 kb (Figure 5).

Agarose gel electrophoresis image demonstrating successful amplification of long cDNA targets generated using Maxima H Minus Reverse Transcriptase in a two-step RT-PCR workflow

Figure 5. Amplification of targets up to 20 kb in two-step RT-PCR. Total RNA was extracted from human and mouse cells and were reverse-transcribed with Maxima H Minus RT (1 µg of total RNA per reaction), human cells (lanes 1 and 2) or mouse cells (lanes 3 and 4). The resulting cDNAs were used as templates for PCR, and the PCR products were visualized by agarose gel electrophoresis. M: Thermo Scientific GeneRuler 1 kb Plus DNA Ladder.

The Maxima H Minus cDNA Synthesis Master Mix shows reliable cDNA synthesis from degraded RNA, maintaining sensitivity and consistent RT-qPCR performance across targets (Figure 6).

In addition, Maxima H Minus RT generated high quality cDNA in the presence of common inhibitors, helping ensure reliable cDNA synthesis and reproducible results across challenging sample types (Figure 7). The common inhibitors include:

  • Contaminants from sample preparation, such as ethanol
  • Plant-derived compounds, such as xylan
  • Blood components, such as heparin

Figure 6. Highest cDNA yield from degraded RNA using Maxima H Minus Reverse Transcriptase. cDNA yield of Maxima H Minus RT was compared to other commercial kits using 100 ng of degraded total HeLa RNA (RIN = 3). Performance in qPCR was evaluated using Applied Biosystems TaqMan Fast Advanced Master Mix and results are shown as ΔCt values [ΔCt = Ct competitor − Ct Maxima H Minus Master Mix]. Maxima H Minus RT offers superior or equivalent performance on challenging RNA samples compared to other commercial RTs.

 

Figure 7. Robust performance in the presence of inhibitors. RT-qPCR was performed using 100 ng of total HeLa RNA containing indicated biological and sample-prep inhibitors, comparing Maxima H Minus RT with competitor kits. qPCR performance was evaluated with Applied Biosystems TaqMan Fast Advanced Master Mix. Maxima H Minus RT showed earliest Ct values across all conditions, indicating strong performance in the presence of inhibitors, while competitor kits exhibited delayed amplification or a complete signal loss (Ct values shown as= 40).

Thermostability

Compared to tested reverse transcriptases, Maxima H minus RT produced higher yields of full-length cDNA over a wide temperature range (Figure 1). Its tolerance to high reaction temperatures allows efficient transcription of RNA regions with extensive secondary structures and helps to improve primer specificity, thereby increasing overall yields.

Agarose gel electrophoresis results comparing cDNA synthesis performance of seven reverse transcription kits.

Figure 1. Reliable performance across temperature range and efficient cDNA synthesis. cDNA was synthesized from Invitrogen Millenium™ RNA Markers by incubating samples at different temperatures for 10 min, using (A) Maxima H Minus Reverse Transcriptase and reverse transcriptases from other suppliers: (B) NEB LunaScript® RT SuperMix Kit, (C) NEB ProtoScript® II First Strand cDNA Synthesis Kit, (D) BioRad Reliance Select cDNA Synthesis Kit, (E) BioRad iScript™ Advanced cDNA Synthesis Kit, (F) Quanta qScript Ultra SuperMix and (G) Quanta qScript Ultra Flex Kit. cDNA products were detected by agarose gel electrophoresis.

Linearity and efficient cDNA synthesis

Maxima H Minus Reverse Transcriptase efficiently synthesizes cDNA from a wide range of input RNA, has higher yields, and better linearity in cDNA synthesis compared to commercially available reverse transcriptases, making it a good choice for RT-qPCR experiments (Figure 2, 3 and 4).

Figure 2. High sensitivity and yield. Maxima H Minus Reverse Transcriptase compared with other commercially available RTs using 5 orders of magnitude (10 pg–1 μg) of total HeLa RNA input in RT-qPCR. qPCR performance was evaluated with Applied Biosystems TaqMan Fast Advanced Master Mix. The results demonstrate strong linearity, with high correlation coefficient (R² = 0.996) and robust amplification efficiency (eff% = 90%). Compared to other commercial reverse transcriptases, Maxima H Minus RT delivers earlier Ct values (red line), indicating higher cDNA yield and detection sensitivity.

Figure 3. Maxima H Minus RT delivers earlier Ct values compared to other commercial RTs. Reverse transcription efficiency of Maxima H Minus RT was compared with commercial RTs by RT-qPCR using 100 ng of total HeLa RNA as input. qPCR performance was evaluated with Applied Biosystems TaqMan Fast Advanced Master Mix and results are shown as ΔCt [ΔCt = Ct competitor − Ct Maxima H Minus RT]. Bars represent the Ct delay observed with competitor kits relative to Maxima H Minus Reverse Transcriptase across multiple gene targets. Lower Ct values obtained with Maxima H Minus indicate efficient cDNA synthesis and consistent sensitivity across the genes tested.

 

four RT-qPCR amplification plots comparing the performance of Maxima H Minus cDNA Synthesis Master Mix with several commercial reverse transcription kits

Figure 4. Enhanced transcription efficiency of Maxima H Minus cDNA Synthesis Master Mix. The Maxima H Minus cDNA Synthesis Master Mix demonstrates better efficiency over a wide range of input RNA amounts when compared to competitor’s RTs. Amplification of the human 18S RNA gene was performed on 10-fold serial dilutions of HeLa total RNA (1 μg to 0.1 pg). First-strand cDNA was generated using the Maxima H Minus cDNA Synthesis Master Mix, Thermo Scientific Maxima First Strand cDNA Synthesis Kit for RT-qPCR, and reverse transcriptases from four other suppliers. cDNA was amplified using the Luminaris Probe qPCR Master Mix, low ROX on the ViiA 7 Real-Time PCR System. Amplification plots indicate variation of ΔRn with cycle number.

Full-length cDNA synthesis

Maxima H Minus Reverse Transcriptase demonstrates up to 50x higher processivity than wild-type M-MuLV RT, enabling efficient synthesis of full-length cDNA from a wide range of RNA templates, including very long transcripts up to 20 kb (Figure 5).

Agarose gel electrophoresis image demonstrating successful amplification of long cDNA targets generated using Maxima H Minus Reverse Transcriptase in a two-step RT-PCR workflow

Figure 5. Amplification of targets up to 20 kb in two-step RT-PCR. Total RNA was extracted from human and mouse cells and were reverse-transcribed with Maxima H Minus RT (1 µg of total RNA per reaction), human cells (lanes 1 and 2) or mouse cells (lanes 3 and 4). The resulting cDNAs were used as templates for PCR, and the PCR products were visualized by agarose gel electrophoresis. M: Thermo Scientific GeneRuler 1 kb Plus DNA Ladder.

Robust cDNA synthesis

The Maxima H Minus cDNA Synthesis Master Mix shows reliable cDNA synthesis from degraded RNA, maintaining sensitivity and consistent RT-qPCR performance across targets (Figure 6).

In addition, Maxima H Minus RT generated high quality cDNA in the presence of common inhibitors, helping ensure reliable cDNA synthesis and reproducible results across challenging sample types (Figure 7). The common inhibitors include:

  • Contaminants from sample preparation, such as ethanol
  • Plant-derived compounds, such as xylan
  • Blood components, such as heparin

Figure 6. Highest cDNA yield from degraded RNA using Maxima H Minus Reverse Transcriptase. cDNA yield of Maxima H Minus RT was compared to other commercial kits using 100 ng of degraded total HeLa RNA (RIN = 3). Performance in qPCR was evaluated using Applied Biosystems TaqMan Fast Advanced Master Mix and results are shown as ΔCt values [ΔCt = Ct competitor − Ct Maxima H Minus Master Mix]. Maxima H Minus RT offers superior or equivalent performance on challenging RNA samples compared to other commercial RTs.

 

Figure 7. Robust performance in the presence of inhibitors. RT-qPCR was performed using 100 ng of total HeLa RNA containing indicated biological and sample-prep inhibitors, comparing Maxima H Minus RT with competitor kits. qPCR performance was evaluated with Applied Biosystems TaqMan Fast Advanced Master Mix. Maxima H Minus RT showed earliest Ct values across all conditions, indicating strong performance in the presence of inhibitors, while competitor kits exhibited delayed amplification or a complete signal loss (Ct values shown as= 40).

Integrated gDNA removal

Some formats of Maxima H Minus Reverse Transcriptase kits and Maxima First Strand cDNA Synthesis Kits are integrated with dsDNA, which facilitates a simplified workflow by combining gDNA removal and cDNA synthesis in a single reaction tube. The dsDNase enables fast, efficient, and complete gDNA removal compared to conventional DNase I treatment (Figure 8). RNA samples treated with dsDNase show no decrease in RNA integrity or quantity (Figure 9). With the use of dsDNase treatment prior to cDNA synthesis, the risk of sample loss can be minimized.

Diagram comparing a traditional reverse transcription workflow with a streamlined Maxima H Minus cDNA synthesis workflow that incorporates integrated dsDNase treatment.
 Click image to enlarge

Figure 8. Integrated dsDNase enables removal of gDNA in 2 minutes.

 

RT-qPCR amplification curves comparing cDNA synthesis reactions performed with and without a dsDNase treatment step

Figure 9. dsDNase treatment does not compromise RNA quality and quantity. Serial dilutions of total RNA were used in cDNA synthesis with Maxima First Strand cDNA Synthesis kits with and without dsDNase step. No changes in Cq values were observed between the compared protocols.

How to synthesize cDNA and eliminate genomic DNA contamination in one tube

Thermo Scientific Maxima First Strand cDNA Synthesis Kits with dsDNase: A complete system for highly efficient synthesis of first strand DNA.

Maxima H Minus Reverse Transcriptase: Citations

Maxima H Minus Reverse Transcriptase is widely used in RNA-seq and single-cell transcriptomics workflows for its excellent performance, sensitivity, and ability to generate long transcripts.

In a comparative study of 11 reverse transcriptases with low-RNA-input conditions, Maxima H Minus RT was identified as one of the top-performing enzymes alongside Invitrogen SuperScript IV Reverse Transcriptase.

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UseReference
​cDNA synthesis step of research into the alternative splicing landscape of infarcted mouse heart identified isoform-level therapeutic targets using state-of-the-art long-read sequencing.Xia B, Shen J, Zhang H et al. (2024) The alternative splicing landscape of infarcted mouse heart identifies isoform level therapeutic targets.Sci Data 11(1):1154.
cDNA synthesis protocol for generating high-quality cDNA for microexon validation and circRNA enrichment.Rahimi K, Venø MT, Dupont DM et al. (2021) Nanopore sequencing of brain-derived full-length circRNAs reveals circRNA-specific exon usage, intron retention and microexons.Nat Commun 12(1):4825.
Optimization of cDNA library preparation based on Switching Mechanism at the 5ʹ end of RNA Transcript (SMART) for ultralow RNA-seq in mice brain tissues.Jia E, Shi H, Wang Y et al. (2021) Optimization of library preparation based on SMART for ultralow RNA-seq in mice brain tissues.BMC Genomics 22:809.
DEPICT-seq for single-cell transcriptomic analysis of rare cell subsets isolated via nucleic acid cytometryBao K, Jiang X, Hu H et al. (2024) DEPICT-seq: Single-cell transcriptomic analysis of rare cell subsets isolated via nucleic acid cytometry.Anal Chem 96(41):16236–16245.
Sensitive and powerful single-cell RNA sequencing using mcSCRB-seqBagnoli JW, Ziegenhain C, Janjic A et al. (2018) Sensitive and powerful single-cell RNA sequencing using mcSCRB-seq.Nat Commun 9:2937.
Simultaneous profiling of RNA isoforms and chromatin accessibility of single cells of human retinal organoidsZhang S, Xiao Y, Mo X et al. (2024) Simultaneous profiling of RNA isoforms and chromatin accessibility of single cells of human retinal organoids.Nat Commun 15:8022.
Simultaneous multiplexed amplicon sequencing and transcriptome profiling in single cells.Saikia M, Burnham P, Keshavjee SH et al. (2019) Simultaneous multiplexed amplicon sequencing and transcriptome profiling in single cells.Nat Methods 16(1):59–62.
Reverse transcription prior to single-cell RNA sequencing (scRNA-seq) of (Saccharomyces cerevisiae and Candida albicans) using microbial Drop-seq technique.Dohn R, Xie B, Back R et al. (2022) mDrop-Seq: Massively parallel single-cell RNA-seq of Saccharomyces cerevisiae and Candida albicans. Vaccines (Basel) 10(1):30.
Droplet-based single-cell RNA sequencing of paraformaldehyde-fixed cellsPhan HV, van Gent M, Drayman N et al. (2021) Droplet-based single-cell RNA sequencing of paraformaldehyde-fixed cells.Protoc Exch.
inDrops-2—a droplet microfluidic approach for high-throughput scRNA-seq of fresh and preserved clinical samplesJuzenas S, Goda K, Kiseliovas V et al. (2025) inDrops-2: A flexible, versatile and cost-efficient droplet microfluidic approach for high-throughput scRNA-seq of fresh and preserved clinical samples. Nucleic Acids Res 53(2):gkae1312.
Hydrop enables droplet-based single-cell ATAC-seq and single-cell RNA-seq using dissolvable hydrogel beadsDe Rop FV, Ismail JN, Bravo González-Blas C et al. (2022) Hydrop enables droplet-based single-cell ATAC-seq and single-cell RNA-seq using dissolvable hydrogel beads.eLife 11:e73971.
High-plex protein and whole transcriptome co-mapping at cellular resolution with spatial CITE-seq.Liu Y, DiStasio M, Su G et al. (2023) High-plex protein and whole transcriptome co-mapping at cellular resolution with spatial CITE-seq.Nat Biotechnol 41(10):1405–1409.
Parallel analysis of individual cells by RNA-seq using drop-seq techniqueMacosko EZ, Basu A, Satija R et al. (2015) Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets.Cell 161(5):1202–1214.

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