FastGene Scriptase II 5x Ready-Mix: Streamlined High-Fidelity cDNA Synthesis
The FastGene Scriptase II 5x Ready-Mix is a comprehensive, all-in-one solution designed to simplify the reverse transcription workflow without compromising on sensitivity or yield. In high-throughput molecular biology environments, the preparation of multiple reaction components can introduce variability and increase the risk of contamination. This 5x Ready-Mix provides an expert solution by combining the engineered enzyme with all necessary buffers, dNTPs, and primers in a single tube, ensuring maximum consistency and reliability for sensitive downstream applications such as RT-qPCR and Next-Generation Sequencing (NGS).
The core of this system is the FastGene Scriptase II, an engineered version of the Murine Leukaemia Virus (MuLV) reverse transcriptase. Strategic mutations within the RNase H domain effectively eliminate endogenous RNase H activity, preventing the degradation of the RNA template during first-strand synthesis. This allows for the production of high-yield, full-length cDNA fragments up to 12 kbp. Additionally, the enzyme features enhanced thermal stability, enabling reactions to be performed at higher temperatures to resolve complex secondary structures and provide a complete picture of gene modifications, including splicing variants.
Key Benefits:
- All-in-One Convenience: Ready-to-use 5x formulation. Contains the enzyme, buffer, dNTPs, RNase inhibitor, and random hexamers, reducing pipetting steps and minimising experimental error.
- Full-Length cDNA Production: Fragments up to 12 kbp. The eliminated RNase H activity prevents template degradation, ensuring the synthesis of long, representative cDNA pools for transcriptome analysis.
- Superior Thermal Stability: Overcome structured templates. Higher operating temperatures allow for the effective transcription of GC-rich and complex RNA that often stalls standard enzymes.
- High Sensitivity for Low-Input: Preserve precious samples. Specifically optimised to produce high-quality cDNA from very low amounts of starting material, ideal for limited clinical or environmental samples.
- Optimised for NGS and qPCR: Validated performance. Delivers superior templates for quantitative assays and sequencing, providing results that compare favourably to the market’s leading brands.
Technical Insights
Figure 1: qPCR Sensitivity Comparison (GAPDH)
Comparison of qPCR results using primers for GAPDH and cDNA produced from varying RNA concentrations. Results demonstrate that FastGene Scriptase II provides nearly identical Ct values to Competitor I’s SS-II enzyme across a broad range of inputs.
Figure 2: qPCR Consistency Analysis (YWHAZ)
Analytical comparison using YWHAZ primers confirms that cDNA derived from Scriptase II maintains high fidelity and quantitative accuracy, matching the performance of premium market standards at 42°C.
Figure 3: Multiplex PCR Performance
Comparison of multiplex PCR results using cDNA produced by Competitor I’s SS-II enzyme and FastGene Scriptase II at both 42°C and 50°C. The data shows that Scriptase II is indistinguishable from leading brands in complex amplification assays.
Specifications
- Catalogue No: P8-0055
- Product: 5x FastGene Scriptase II cDNA Synthesis Kit Ready-Mix
- Format: 5x Concentrated Ready-Mix (100 Reactions)
- Enzyme Type: Engineered MuLV Reverse Transcriptase (RNase H Minus)
- Included Components: Scriptase II Enzyme, 5x Buffer, dNTPs, RNase Inhibitor, Random Hexamers, Helper Protein
- Max Fragment Size: 12 kbp
- Recommended Applications: RT-qPCR, NGS Library Prep, Multiplex PCR
- Storage: Store at -20°C for long-term stability