FastGene Scriptase II - Ready Mix OdT (including oligo dT & random hexamers)

Ready Mix - Ready-to-use including random hexamers and Oligo-dTs

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 revers 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.
 
Applications

• Quantification of Gene Expression
• RT-qPCR
• Next Generation Sequencing
• Low RNA concentration
• Difficult templates

 
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.

Composition of mix:

• FastGene® Scriptase II
• 5x FastGene® Scriptase II buffer
• dNTP Mixture
• RNase Inhibitor
• Random Hexamer
• oligo dTs
• Helper Protein

Protocol

1. Mix the ReadyMix with the RNA Template using the following setup:

5x FastGene® Scriptase II ReadyMix (LS64)    4 µl
High quality RNA-template*                                x µl
Sterile Water                                           up to 20 µl

*Concentration of template RNA can be up to 2.5 μg

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.