FastGene Scriptase III cDNA Synthesis Enzyme: Thermostable Precision for High-Yield Reverse Transcription
The FastGene Scriptase III Synthesis Enzyme is the ideal choice for researchers requiring maximum flexibility in their experimental design. Provided as a high-concentration standalone enzyme with a dedicated 5X buffer, this format allows for precise control over reaction conditions, including the choice of specialised primers and additive concentrations. It is specifically engineered to handle complex RNA templates, maintaining high activity at temperatures up to 55°C to ensure the synthesis of full-length cDNA even from extensively structured targets.
The FastGene Scriptase III range provides a sophisticated solution for researchers requiring highly efficient and reliable reverse transcription, particularly when dealing with challenging RNA templates. Standard M-MLV reverse transcriptases often struggle with complex secondary structures and low-input samples, leading to truncated cDNA or biased results. FastGene Scriptase III overcomes these limitations, offering an engineered alternative that ensures high-fidelity synthesis of full-length cDNA across a wide range of applications, including qPCR, NGS, and cDNA library construction.
At the core of this technology is an advanced, engineered variant of the Murine Leukaemia Virus (MuLV) reverse transcriptase. By incorporating specific mutations within the RNase H domain, the enzyme’s degradative activity is significantly reduced, allowing for the synthesis of long cDNA transcripts up to 12 kb. Furthermore, additional stabilising mutations provide exceptional thermostability, enabling the enzyme to remain fully active at temperatures up to 55°C. This elevated operating temperature is critical for unwinding extensively structured RNA, ensuring the entire template is accessible for transcription and resulting in superior sensitivity and yield.
Key Benefits:
- Enhanced Thermostability: Superior performance at 55°C. Operates effectively at higher temperatures to resolve complex RNA secondary structures, ensuring complete template coverage.
- Reduced RNase H Activity: Synthesise long cDNA transcripts. The engineered RNase H domain prevents premature template degradation, facilitating the production of cDNA up to 12 kb in length.
- Exceptional Sensitivity: Reliable detection from 0.1 ng RNA. Optimised for low-input samples, providing consistent and reproducible results even with minimal starting material.
- High-Speed Protocols: Rapid results in just 30 minutes. The high processivity of the enzyme allows for significantly shortened incubation times without compromising on yield or quality.
- Flexible Formats: Tailored to your workflow. Available as a standalone enzyme, a complete cDNA synthesis kit with primers, or a convenient 5x ReadyMix for maximum throughput.
Technical Insights
▼ Technical Performance Data & Protocols
Boost your cDNA synthesis with thermostable precision
Scriptase III sets a new standard for reverse transcription with its enhanced thermostability, allowing efficient operation at temperatures up to ~55°C. This elevated temperature helps unwind RNA secondary structures, granting better accessibility and increasing the sensitivity of reverse transcription reactions. The result? Higher yields of full-length, high-quality cDNA.
Figure 1: Thermostability Performance
Performance of the Scriptase III at different temperatures. The thermostability of the Scriptase III ensures a reliable operation up to ~55°C.
Amplify productivity with superior cDNA yields
Boost your productivity with Scriptase III. Real-time PCR analysis reveals significantly lower Cq values, indicating its efficiency in generating abundant and high-quality cDNA even from minimal RNA input.
Figure 2: Yield Comparison
Productivity of Scriptase III. Scriptase III shows excellent productivity and cDNA yields for different RNA targets, compared to competitor enzymes.
Incredible sensitivity
Achieve precise cDNA synthesis even from minimal RNA samples with Scriptase III. This advanced reverse transcription enzyme ensures reliable detection down to 0.1 ng of total RNA, as demonstrated with HeLa RNA extracts. Real-time PCR results show consistent GAPDH amplification across RNA concentrations ranging from 100 ng to 0.1 ng, showing Scriptase III’s unparalleled sensitivity and efficiency. Perfect for low-input applications, Scriptase III delivers consistent, high-quality cDNA to support your most demanding research needs.
Figure 3: Sensitivity Analysis
Sensitivity of Scriptase III. Scriptase III ensures reliable detection down to 0.1 ng of total RNA extracts.
High-speed cDNA synthesis
When time is critical, Scriptase III rises to the challenge. Its reaction time can be shortened to 30 minutes if needed, without sacrificing cDNA yield or performance. This flexibility makes it an excellent choice for high-quality results even under a tight schedule.
Figure 4: Reaction Time Efficiency
Reaction time for Scriptase III. Fast reaction times of Scriptase III allow a reduction of the incubation time down to 30 minutes.
Product Variations
| Catalogue No. | Product | Contents/Specifications |
|---|---|---|
| P8-0051 | FastGene Scriptase III Synthesis Enzyme | Includes Scriptase III Enzyme (200 U/µL), 5X First-Strand Buffer, 0.1 M DTT, dNTP mix (10 mM each), and RNase-free water. Primers not included. |
| P8-0049 | FastGene Scriptase III cDNA Kit | Complete kit for 100 reactions. Includes Enzyme, 10X RT Buffer, MgCl2, DTT, dNTPs, RNase Inhibitor, Oligo dT, and Random Hexamers. |
| P8-0053 | FastGene Scriptase III cDNA 5x ReadyMix | Convenient 5x concentrated master mix containing all components (except template and water) for high-throughput cDNA synthesis workflows. |
Specifications
- Reaction Temperature: Optimised for 42°C to 55°C.
- Transcript Length: Up to 12 kb.
- Input Sensitivity: Down to 0.1 ng total RNA.
- RNase H Activity: Minimised for high-yield full-length cDNA.
- Storage Conditions: Store at -20°C in a non-frost-free freezer.
- Shelf Life: 12 months when stored correctly.
- Applications: RT-qPCR, NGS, cDNA Library Construction, RNA Analysis.