FastGene® Scriptase II cDNA 5x ReadyMix
The FastGene® Scriptase II 5x ReadyMix contains an engineered enzyme with a reduced RNase H activity and comes ready-to-use with all necessary ingredients to perform a revers transcription.
- Ready-to-use with all necessary ingredients to perform a revers transcription.
- Drop-in replacement for Superscript II
check out comparison data
Applications
- Quantification of Gene Expression
- RT-qPCR
- Next Generation Sequencing
- Low RNA concentration
- Difficult templates
| Cat-No. | Item | Size | Price (CHF) |
|---|---|---|---|
| LS65 | FastGene Scriptase II cDNA 5x ReadyMix (100 rxns) (incl. Oligo dT and randon hexamers) | 100 reactions | 415.00 |
Composition
The FastGene® Scriptase II 5x ReadyMix comes with all necessary ingredients to perform a revers transcription:
- 5x Buffer
- dNTP Mixture
- RNase Inhibitor
- Random Hexamer
- Helper Protein
Details
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 (LS64) 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.
Protocol
1. Mix the ReadyMix with the RNA Template using the following setup:
| 5x FastGene® Scriptase II ReadyMix (LS65) | 4 µL |
| High quality RNA-template (concentration of template RNA can be up to 2.5 μg) | x µL |
| Sterile Water | up to 20 µL |
2. Incubate at 25 °C for 10 minutes.
3. Incubate at 42 °C for 60 minutes.
- Note: The recommended incubation time can be reduced to 5 minutes depending on the product abundance and size. Download here our Technical Note showing the performance at lower temperature (down to 5 min.).
4. Inactivate the FastGene Scriptase II by incubating at 85°C for 5 min.
5. Store the cDNA at -20 °C or use it for downstream application.