FastGene Scriptase II cDNA Synthesis Kit: Engineered Precision for Complex RNA Templates
The FastGene Scriptase II cDNA Synthesis Kit is a sophisticated solution developed to overcome the inherent challenges of reverse transcription, particularly when dealing with low-abundance or highly structured RNA. In standard cDNA synthesis, the endogenous RNase H activity of reverse transcriptases can degrade the RNA template before the first strand is fully completed, leading to truncated products and reduced library representation. This kit provides a professional-grade alternative, ensuring the synthesis of high-fidelity, full-length cDNA even from the most demanding starting materials.
The core of this kit is an engineered version of the Murine Leukaemia Virus (MuLV) reverse transcriptase, featuring strategic mutations within the RNase H domain. By eliminating RNase H activity, the enzyme allows the RNA-DNA heteroduplex to remain stable throughout the entire synthesis process, resulting in significantly higher yields of full-length transcripts. Furthermore, the enzyme has been modified for enhanced thermal stability, allowing the reaction to be performed at elevated temperatures. This is critical for melting complex secondary structures in GC-rich templates, making it the optimal engine for sensitive downstream applications such as RT-qPCR and Next-Generation Sequencing (NGS) library preparation.
Key Benefits:
- Zero RNase H Activity: Maximise full-length cDNA yields. The elimination of RNase H prevents the degradation of the original RNA template during synthesis, ensuring complete transcript representation.
- Superior Thermal Stability: Overcome secondary structures. High-temperature compatibility allows for the effective transcription of GC-rich and highly structured RNA that often stalls standard enzymes.
- Comprehensive Kit Format: Complete synthesis solution. Supplied with Oligo dTs, random hexamers, and a high-potency RNase inhibitor to provide a seamless and protected workflow.
- High Sensitivity: Ideal for low-input samples. Specifically optimised to produce high-quality cDNA from very low amounts of starting material, preserving precious clinical or environmental samples.
- Optimised for NGS & qPCR: Reliable downstream performance. The combination of high fidelity and robustness ensures consistent results in high-throughput sequencing and quantitative assays.
Technical Insights
Impact of RNase H Mutation on cDNA Length
The engineering of the RNase H domain is the defining feature of FastGene Scriptase II. In traditional enzymes, the RNase H activity cleaves the RNA template during synthesis, which often results in the loss of the 5′ ends of long transcripts. By removing this activity, Scriptase II maintains the integrity of the RNA-DNA hybrid, allowing the polymerase to reach the end of the template. This results in a more representative cDNA pool, which is a critical requirement for accurate gene expression profiling and transcriptome sequencing.
Specifications
- Enzyme Type: Engineered MuLV Reverse Transcriptase (RNase H Minus)
- Thermal Stability: High-temperature compatible
- Kit Components: Scriptase II Enzyme, Oligo dTs, Random Hexamers, RNase Inhibitor
- Recommended Applications: RT-qPCR, NGS Library Prep, Complex/GC-rich RNA
- Storage: Store at -20°C for long-term stability