Meridian is a primary manufacturer of specialized high-quality molecular reagents and offers solutions to a wide range of industries to diagnose and treat diseases, discover new therapeutics or develop tests for environmental, food and cosmetic safety.
Glycerol-Free T7 RNA Polymerase High concentration (1,000 U/ µ L),
glycerol-free and ideal for large-scale RNA synthesis
Glycerol-Free T7 RNA Polymerase (HC) is a high concentration (1,000 U/ μ L) DNA-dependent RNA Polymerase with very high specificity for the T7 phage double- stranded promoter sequence. The enzyme catalyzes the 5’ → 3’ synthesis of RNA on either single-stranded DNA or double-stranded DNA downstream from the promoter. T7 RNA Polymerase is a DNA-dependant RNA Polymerase originating from the T7 bacteriophage and is the industry standard enzyme for in vitro transcription (IVT). It can efficiently incorporate standard or modified ribonucleoside triphosphates (NTPs) synthesizing large amounts of mRNA very quickly, making it an ideal enzyme for in vitro synthesis of labeled or unlabeled RNA for research, diagnostic or clinical applications. Applications including isothermal diagnostic methods, such as nucleic acid sequence-based amplification (NASBA), mRNA vaccine production, and RNA-based therapeutic and research applications, including miRNA and siRNA synthesis for RNA interference drugs, synthesis of sgRNA for the CRISPR/Cas9 system. Glycerol-Free T7 RNA Polymerase (HC) offers superior performance without the presence of glycerol, allowing the development of reliable in vitro transcription workflows. Additionally, it provides the added capability of lyophilization, granting users the flexibility
to store reaction mixtures at room temperature or produce diagnostic assays with miniaturized reaction components.
Product applications
PRODUCT
CAT NO. VOLUME REACTIONS
• Synthesis of RNA (including mRNA and sgRNA)
50 uL 50,000 Units
Glycerol-Free T7 RNA Polymerase (HC)
MDX201
• Synthesis of RNA probes for hybridization or RNA templates for in vitro RNA translation • Radiolabelled/fluorescently labelled RNA probe preparation • Generation of antisense RNA • Nucleic acid sequence-based amplification (NASBA)
500 uL 500,000 Units
www.meridianbioscience.com/lifescience
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.
Ordering information: USA 5171 Wilfong Road Memphis, Tennessee 38134 Phone: +1 901-382-8716 Fax: +1 901-333-8223
Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.meridianbioscience.com/lifescience
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