Engineered M‑MuLV for first‑strand cDNA synthesis
Engineered M‑MuLV RT with reduced RNase H activity and increased thermostability supports reverse transcription up to 55 °C. Generate long cDNA and start RNA‑seq libraries with reliable performance on structured RNAs.
| Product Number | EM16EMMULV |
| Molecular Weight | 77 kDa |
| Function | RNA-dependent DNA polymerase with reduced RNase H |
| For Use In | cDNA synthesis; RT-PCR; Library initiation |
| Concentration | 200 U/µL |
| Delivery Format | Tubes |
| Intended Use | This product is intended for research use only |
| Volume | 100 µL 500 µL |
| Source | Recombinant E. coli |
| Shipping Conditions | Cold Pack |
| Storage Temperature | -20°C |
| Stability | ≥12 months |
Synthego’s Reverse Transcriptase is a genetically engineered M-MuLV Reverse Transcriptase designed for exceptional performance in cDNA synthesis and RT-PCR applications. With increased thermostability and reduced RNase H activity, our engineered M-MuLV delivers high yields of full-length cDNA, even for longer templates, making it a reliable choice for demanding workflows.
This ultrapure enzyme is free of RNases and nucleases, ensuring optimal reaction conditions and preventing degradation of RNA templates. It is active at temperatures up to 55°C, enabling first-strand cDNA synthesis at higher temperatures, which improves specificity and reduces secondary structure interference. It is ideal for both two-step and one-step RT-PCR and qRT-PCR, offering superb sensitivity and specificity, even with low template inputs.
We offer flexible customization to meet your specific needs:
One unit is the amount of enzyme activity that incorporates 1 nmol of dTTP into acid insoluble fraction in 10 minutes at 42°C when poly(A)+ RNA and oligo(dT)20 are used as template–primer.
Exonuclease assay
Linearized lambda/HindIII fragments are incubated with the Reverse Transcriptase in a 50 µl reaction mixture for 4 hours at 37°C. No degradation of DNA was observed.
Endonuclease/Nick Activity
Supercoiled plasmid DNA is incubated with the enzyme in a 50 µl reaction mixture for 4 hours at 37°C. No conversion of covalently closed circular DNA to nicked DNA detected.
Contamination with E. coli DNA
Absence of E. coli genomic DNA is confirmed by qPCR using a sample of the enzyme and specific primers targeting the E. coli 16S rRNA gene. No contamination detected.
RNase Assay
An RNA template is incubated with the enzyme in a 20 µl reaction mixture for 1 hour at 42°C. No RNA degradation observed.
Functional Assay
cDNA synthesis with oligo(dT) and/or hexamer primers, followed by PCR.
Common primer options include:
- Oligo(dT) primers: Target poly(A) tails of mRNA and enrich for mature transcripts
- Random primers: Enable broad cDNA synthesis from various RNA species
- Gene-specific primers: Provide targeted reverse transcription of specific RNA transcripts
The recommended RNA input depends on the reverse transcription protocol and downstream application. We recommend using at least 50 ng of RNA template in your reactions.
High-quality template RNA is one of the most important factors for maximizing cDNA yield. Include Synthego RNase Inhibitor in your RNA isolation, optimize primers used for your reverse transcription, and ensure appropriate reaction conditions.
Yes. The workflow described is a two-step RT-PCR process, where cDNA is synthesized in an initial reaction and the resulting product is transferred directly—without prior purification—into a separate downstream PCR. Up to 10 µL of the finished cDNA mix can be used per 50 µL qPCR volume.
While this FAQ addresses two-step workflows, the Engineered M-MuLV Reverse Transcriptase enzyme itself is versatile and formulated for both one-step and two-step RT-PCR/qRT-PCR:
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