Product Overview

WS HHRevScript™ Reverse Transcriptase is a MMLV-based reverse transcription enzyme designed for use in IVD molecular diagnostics and RT-qPCR assay development. It is engineered to support efficient cDNA synthesis under challenging conditions, including low RNA input, elevated reaction temperatures, and complex sample matrices.

Based on validated experimental data, this enzyme demonstrates improved performance in terms of reaction speed, thermal stability, sensitivity, inhibitor tolerance, and formulation compatibility, making it suitable for a wide range of molecular diagnostic workflows.

WS HHRevScript™ Reverse Transcriptase is compatible with:

• RT-qPCR assays

• One-step RT-PCR systems

• Direct amplification workflows (with lysis reagents)

• Pre-mixed diagnostic reagent formulations

Key Features

Fast cDNA Synthesis (2-minute RT reaction)

HHRevScript™ Reverse Transcriptase enables rapid reverse transcription.
Experimental results show that a 2-minute RT reaction achieves comparable qPCR sensitivity to a 10-minute reaction under identical PCR conditions.

High Thermal Stability

The enzyme maintains stable performance under elevated temperatures.

• No significant loss of activity after incubation at 55°C for 30 minutes

• Retains performance in RT-qPCR reactions conducted at 55–60°C

• Supports reverse transcription under high-temperature conditions

High Sensitivity for Low RNA Input

The enzyme demonstrates strong performance across a wide RNA input range.

• Tested with 0.1 pg–1 μg HeLa total RNA (6 gradients)

• Achieved detection sensitivity down to 1 pg RNA

• Maintains consistent amplification efficiency across dilution series

Strong Tolerance to PCR Inhibitors

HHRevScript™ Reverse Transcriptase maintains performance in the presence of common inhibitors, including:

• Freshly prepared nucleic acid release reagent

• Swab sample matrix

• 10 ng/L humic acid

• 400 μM hemoglobin

• 20 μM heme

• 80 μg/mL mucin

ΔCt analysis shows improved robustness under inhibitor-containing conditions compared with standard reactions.

Compatibility with Direct Amplification Systems

The enzyme performs reliably in workflows using crude sample input.

• Compatible with nucleic acid release reagents

• Supports direct amplification from swab-based samples

• No significant performance loss observed in multiplex RT-qPCR systems with lysis reagents

Stable Performance in Pre-mixed Formulations

HHRevScript™ Reverse Transcriptase demonstrates formulation stability in fully pre-mixed systems.

• Compatible with reaction buffer, enzyme, and primers/probes pre-mixed in one tube

• Stability tested at 2–8°C for 14 days and 37°C for 7 days

• No significant performance difference compared with freshly prepared controls under a 30-minute rapid PCR protocol

Applications

WS HHRevScript™ Reverse Transcriptase is suitable for:

• RT-qPCR-based pathogen detection

• RNA virus molecular diagnostics

• Clinical molecular testing workflows

• Rapid diagnostic assay development

• Direct amplification systems using lysis reagents

• Pre-mixed IVD reagent kit development

Performance Data

Ultra-Fast cDNA Synthesis

HHRevScript™ Reverse Transcriptase III has been engineered to increase the rate of cDNA synthesis. Under identical PCR conditions, a 2-minute reverse transcription step achieves comparable sensitivity to a 10-minute reaction, demonstrating significantly improved reaction efficiency without compromising assay performance.

Enzyme Thermal Stability (Residual Activity Assay)

Different reverse transcriptases were incubated at 50°C–60°C for 30 minutes, and residual enzyme activity was measured.

The results show that HHRevScript™ Reverse Transcriptase III:

• Maintains full activity after incubation at 55°C for 30 minutes

• Exhibits a half-life longer than 30 minutes at 60°C

These results demonstrate strong thermal stability under elevated temperature conditions.

RT-qPCR Performance at Elevated Temperatures

Different reverse transcriptases were evaluated in RT-qPCR reactions performed at 45°C, 50°C, 55°C, and 60°C.

The results demonstrate that HHRevScript™ Reverse Transcriptase III:

• Maintains stable reverse transcription performance at 55°C–60°C

• Shows no significant decline in amplification efficiency under high-temperature conditions

Sensitivity Across RNA Input Range

HeLa total RNA (0.1 pg–1 μg, 6 serial dilutions) was used as the template for reverse transcription. Subsequently, 1 μL of cDNA was used for qPCR analysis.

The results indicate that HHRevScript™ Reverse Transcriptase III:

• Provides consistent amplification across a wide RNA input range

• Achieves a detection sensitivity of 1 pg RNA

Inhibitor Resistance in RT-qPCR

Reverse transcription was performed using 0.1 μg HeLa total RNA with different reverse transcriptases in the presence of various inhibitors.

Inhibitors included:

• Hemoglobin

• Heme

• Mucin

• Humic acid

After reverse transcription, 1 μL of cDNA was used for qPCR analysis. ΔCt values were compared, where lower ΔCt indicates stronger inhibitor tolerance.

The results show that HHRevScript™ Reverse Transcriptase III exhibits strong resistance to common RT-qPCR inhibitors, demonstrating robust performance in direct amplification workflows.

Direct Amplification Compatibility

In a multiplex RT-qPCR system, nucleic acid release reagents were added, or swab samples were directly used for amplification without purification.

The results demonstrate that HHRevScript™ Reverse Transcriptase III:

• Maintains stable performance in the presence of nucleic acid release reagents

• Is compatible with direct amplification from swab-based samples

• Shows no observable reduction in amplification performance

Pre-mixed Formulation Stability

WSH-CH HHRevScript™ Reverse Transcriptase II was incorporated into a fully pre-mixed system containing reaction buffer, enzyme solution, and multiplex primers/probes in a single tube.

Stability testing was performed under:

• 2–8°C for 14 days

• 37°C for 7 days (accelerated stability condition)

Performance was evaluated using a 30-minute rapid cycling protocol, with freshly prepared formulations as controls.

The results show that the pre-mixed system containing HHRevScript™ Reverse Transcriptase II:

• Exhibits no significant performance difference compared with freshly prepared controls

• Demonstrates excellent formulation stability under stress conditions

FAQ

What are the main features of WS HHRevScript Reverse Transcriptase?

WS HHRevScript Reverse Transcriptase is a high-efficiency reverse transcription enzyme designed for robust cDNA synthesis. It exhibits strong RNA affinity, high extension efficiency, and improved tolerance to common inhibitors, making it suitable for challenging RNA templates and diagnostic workflows.

What types of RNA templates is WS HHRevScript suitable for?

It is suitable for a wide range of RNA templates, including total RNA, viral RNA, and partially degraded RNA samples. It performs well with low-abundance targets and complex clinical specimens.

What is the temperature range and reaction characteristics of this reverse transcriptase?

WS HHRevScript Reverse Transcriptase supports a relatively wide working temperature range and maintains stable activity at elevated temperatures, which helps reduce RNA secondary structure and improves full-length cDNA synthesis efficiency.

Does WS HHRevScript Reverse Transcriptase have RNase H activity?

WS HHRevScript is engineered with reduced or controlled RNase H activity, enabling higher cDNA yield and improved integrity of RNA–cDNA hybrids during reverse transcription.

Can WS HHRevScript be used in one-step RT-qPCR systems?

Yes. It is compatible with one-step RT-qPCR workflows and can be combined with hot-start Taq polymerases and optimized buffer systems for efficient reverse transcription and real-time amplification in a single tube.

What types of inhibitors can WS HHRevScript tolerate?

The enzyme shows improved tolerance to common RT inhibitors such as salts, heme compounds, mucosal components, and other contaminants often present in crude or minimally processed clinical samples.

What downstream applications is WS HHRevScript suitable for?

What is the recommended storage condition and stability?

WS HHRevScript Reverse Transcriptase should typically be stored at -20°C. It remains stable under standard freeze–thaw handling conditions when used with optimized storage buffers and stabilizers.

Is WS HHRevScript compatible with multiplex or low-copy detection systems?

Yes. It provides strong sensitivity and uniform cDNA synthesis performance, making it suitable for multiplex assays and low-copy RNA detection in diagnostic applications.