FastGene® Scriptase Basic
Optimized for better performance
The FastGene® Scriptase Basic is an enhanced version of the Murine Leukemia Virus (MuLV) reverse transcriptase. Hence, like the wildtype, it has the ability to synthesize a cDNA strand, a reduced RNase H activity and processitivity. The robustness however was greatly increased. It is the perfect enzyme for large RNA quantities and easy templates.
- Quantification of Gene Expression
- Endpoint RT-PCR
- High RNA concentrations
No inhibition – Even at large concentration
The special buffer formulation permits a high RNA concentration. Other reverse transcriptases are not able to process such large quantities.
Designed for Endpoint RT-PCR
The FastGene® Scriptase Basic was designed for large RNA quantities, typically used in an endpoint RT-PCR. Nonetheless, it is also able to process lower RNA concentrations.
Enzyme only or cDNA Synthesis kit
The FastGene® Scriptase Basic is available as enzyme only, containing the enzyme, buffer and dNTPs as well as a cDNA synthesis kit which contains the components of the enzyme plus Oligo dTs, random hexamers and RNase inhibitor. We have also chosen a 100 reactions kit size so that you have enough for a whole 96-well plate.
Comparison of FastGene Scriptase Basic and a conventional MuLV
(A) Competitor N (B) FastGene® Scriptase Basic
RNA (ng) 2 1 0.5 0.1 0.01 2 1 0.5 0.1 0.01
Fig. 1: Higher sensitivity of the FastGene® Scriptase Basic.
Compared to a wildtype MuLV (A) the FastGene® Scriptase Basic (B) is an optimized MuLV-Reverse transcriptase which has a higher sensitivity, being able to produce a template from RNA concentrations as low as 0.1 ng.
LS-52-Scriptase Basic Enzyme Manual
Protocol for the FastGene Scriptase Basic
I am using your Scriptase Basic (LS52) and I wanted to know if I can use this enzyme in combination with a DNA removal kit which contains a DNase Stop buffer with 20 mM EGTA?
All DNA/RNA working enzymes need metal ions like Mg2+, Ca2+, Mn2+ and so on. DNase I requires Mg2+ or Ca2+ as well. Therefore, DNase I Stop buffer must contain 20 mM of EGTA or EDTA (as metal chelating agent) to trap the metal ions. For most enzymes the working condition is around 1~10 mM. So, 20 mM of EGTA is enough to stop both reactions, DNase treatment and RT.
To keep Scriptase Basic (LS52) working further on, the concentration of Mg2+ has to be increased to make the final Mg2+ concentration back to the working concentration of the enzyme. But in this case the DNase I will start again and degrade DNA/cDNA.
This is a common problem with all Reverse Transcriptases.
What is the optimal temperature range for Scriptase Basic and Scriptase II?
The optimal temperature range for the Scriptases is 42°C to 50°C. A temperature above 50°C is not recommended.