Product Highlights RNA Yield: gel electrophoresis
Glycerol-Free T7 RNA Polymerase (HC) (MDX201) was lyophilized with Lyo-Ready TM Transcription Buffer. This was tested by performing in vitro transcription reactions against fresh (wet) MDX201, NEB T7 RNA Polymerase and ThermoFisher T7 RNA Polymerase using their recommended reaction conditions. Reactions were incubated at 37°C for up to 24 hours and purified using a column-based RNA purification kit. Purified RNA was analysed on a 1.5% agarose gel, stained with ethidium bromide and visualized by UV fluorescence. These results demonstrate that a considerably higher yield of RNA was synthesized using MDX201 compared to other manufacturers’ kits.
RNA Yield: spectrophotometer analysis
In vitro transcription reactions were assembled, according to the manufacturer’s recommended protocol, using 0.1 ng of dsDNA template encoding a 1 kb or 4.5 kb RNA. The reactions were incubated at 37°C for up to 8 hours with lyophilized Glycerol-Free T7 RNA Polymerase (HC) (MDX201) ( red ), NEB T7 RNA Polymerase ( orange ) and ThermoFisher T7 RNA Polymerase ( black ). These samples were tested by performing in vitro transcription reactions against MDX201 before lyophilization, NEB T7 RNA Polymerase and ThermoFisher T7 RNA Polymerase. At each time point, the corresponding samples were transferred to -20°C to stop the reaction. Transcription reactions were purified using a column-based RNA purification kit after the last time point and quantified on NanoDrop TM Spectrophotometer. These results demonstrate that a considerably higher yield of RNA was synthetized using MDX201.
Enzyme Retains Full Activity After Lyophilization
Glycerol-Free T7 RNA Polymerase (HC) (MDX201) was lyophilized in Lyo-Ready TM Transcription Buffer and subsequently stored at 37°C for accelerated stability studies. These enzymes were tested by performing in vitro transcription reactions against fresh liquid MDX201 stored at -20°C. Reactions were incubated at 37°C for 2 hours and purified using a column-based RNA purification kit. Purified RNA was run on a 1.5% agarose gel, stained with ethidium bromide and visualized by UV fluorescence. These results demonstrate that Glycerol-Free T7 RNA Polymerase (HC) is not affected by lyophilization and retains its activity when rehydrated.
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