Details
ReadyMix
The FastGene® Scriptase II 5x ReadyMix is an engineered enzyme with a reduced RNase H activity and comes ready-to-use with all necessary ingredients to perform a reverse transcription.
Engineered enzyme
Engineered reverse transcriptases allowed the synthesis of cDNA from very low amounts of RNA. Mutations are inserted into the RNase H domain of the MuLV‘s reverse transcriptase. Therefore, by not degrading the RNA during the first-strand synthesis, a higher yield of full-length cDNA is obtained. Additionally, a higher thermal stability increases the robustness of the enzyme. The FastGene® Scriptase II is exactly one of those engineered enzymes. With its mutation in the RNase H domain and higher thermal stability, it is the optimal choice for more complex applications, such as RT-qPCR and NGS.
Lower RNase H activity for longer cDNA
The FastGene® Scriptase II has a modified RNase H domain. The RNA is therefore not degraded and serves as a template for longer cDNAs, resulting in fragment size of up to 12 kBp.
Engineered enzymes – optimized for qPCR
The FastGene® Scriptase II delivers superior cDNA templates for downstream applications, e.g. qPCR and NGS. The resulting full-length cDNA gives a complete picture of the gene and is able to show modification, e.g. splicing variants.
Greatest performance − Product comparison
A current trend in RT enzyme engineering is to withstand very high temperatures while performing a reverse transcription (50+ °C). However, our FastGene® Scriptase II shows a much higher performance even at 42 °C. At that low temperature, the FastGene® Scriptase II can outperform even high-temp enzymes. Have a look at the comparison data with competitor A, which was performed at 55 °C:
Figure: Comparison of the performance of FastGene® Scriptase II (cyan) and a heat-stable reverse transcriptase from Competitor A (orange). FastGene® Scriptase II ReadyMix and Competitor A´s reverse transcriptase were incubated for 5 to 50 minutes at 42 °C or 55 °C, according to the TDS recommendations. FastGene® Scriptase II ReadyMix outperforms the high-temperature enzyme at every gene and time point.
Tsintari Vasileia, Universitätsklinikum Tübingen, Labor für Experimentelle Hämatologie und Onkologie –
Wir haben das Produkt ausprobiert und wir waren mehr als zufrieden damit! Wir haben verschiedene qPCRs durchgeführt mit zwei unterschiedlichen Kontrollen (GAPDH und Tubulin-b), sowohl für Zellinien, als auch für Patienten Proben (Biobank tiefgefrorenen Biopsiegewebe), und die Ergebnisse, waren ganz gut! Die Kontrollwerte sind stabil geblieben im Vergleich zu unserem alten cDNA Protokoll und sogar mit weniger RNA als früher. Anwendung und Protokoll fanden wir ebenso super easy!
Translation by Nippon Genetics:
We use the Sciptase II Ready Mix and we were more than happy with it! We performed different qPCRs with two different controls (GAPDH and tubulin-b), for both cell lines and patient samples (Biobank frozen biopsy tissue). The results were quite good! The control values were stable compared to our old cDNA protocol, even with less RNA than before. The application and the protocol were very easy!
Verified User