- Preparation of Extensively Labeled RNA Probes
- Preparation of Precursors for RNA Splicing Reactions
- Production of Capped mRNA with a Cap Analog as the Primer
- Preparation of mRNA for In Vitro Translation
1.Comparison of Thermostability
We compared thermostability between native T7 RNA Polymerase and this enzyme. Remaining enzymatic activities were measured after incubation of Native T7 RNA Polymerase at 48°C and Thermo T7 RNA Polymerase at 50°C for each reaction time. Results showed that activity decreased to approximately 1/10 after 5-minute incubation at 48°C for the former, whereas activity had barely decreased even after 15-minute incubation at 50°C with Thermo T7 RNA Polymerase.
Half-life at 50°C
Native T7 RNA Polymerase<1.9min.
Thermo T7 RNA Polymerase 84.5min.
2. In Vitro transcription Reaction at High Temperature
We compared in vitro transcription using Native T7 RNA Polymerase and Thermo T7 RNA Polymerase.
20 U of RNase Inhibitor and 25 U of both enzymes were added using 0.5g of plasmid containing the T7 promoter sequence as a template to incubate at 37-51°C for 60 minutes, followed by electrophoresis of reaction products. Results showed that Thermo T7 RNA Polymerase allows transcription even at 51°C.