HHRevScript Reverse Transcriptase is an RNA-directed DNA polymerase capable of synthesizing complementary DNA (cDNA) from either RNA or single-stranded DNA templates, using a primer to initiate synthesis. This enzyme lacks 3′→5′ exonuclease activity and RNase H activity, ensuring high-fidelity reverse transcription. It is thermostable up to 50°C, making it ideal for reverse transcription of RNA templates with complex secondary structures, enhancing binding affinity and reaction efficiency.

One unit incorporates 1 nmol of dTTP into acid-precipitable material in 10 minutes at 37°C using poly(A)•oligo(dT) as template-primer.

Template RNA optional
Oligo(dT)18 (50 μM) or Random Primer Mix (50 ng/μL): 1 μL
dNTP Mixture (10 mM each): 1 μL
RNase Inhibitor (40 U/μL): 1 μL
HHRevScript Reverse Transcriptase (200 U/μL): 1 μL
5× HHRevScript Buffer: 4 μL
Nuclease-free water: Up to 20 μL

25°C 5 min
50°C 45 min
85°C 2 min
If Random Primer Mix is used, include a 25°C incubation step.
For templates with complex secondary structure or high GC content, reaction temperature can be raised to 55°C.
Extended transcription time yields longer cDNA (>5 kb)

To avoid RNase contamination, keep the experimental area clean and wear gloves and masks. Use RNase-free tubes and pipette tips.
The cDNA product can be stored at -20°C or -80°C or used immediately in PCR/qPCR.
Recommended cDNA input volume for PCR/qPCR should not exceed 1/10 of total reaction volume.

Protein purity (SDS-PAGE): ≥95%
Endonuclease activity: No degradation of 1 μg λDNA after 16 hrs at 37°C with 200 U
Exonuclease activity: No degradation of λ-Hind III digest DNA after 16 hrs at 37°C with 200 U
RNase activity: No degradation of MS2 RNA (1.6 μg) after 4 hrs at 37°C with 200 U
E. coli DNA: ≤10 copies/600 U by TaqMan qPCR

Ship with ice packs. Store at -20°C. Avoid repeated freeze-thaw cycles.