T7 P&L RNA Polymerase HC

SKU: orb692276

Description

Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA poly-merase (99 kDa) that catalyzes in vitro RNA synthesis from a DNA sequence containing a T7 phage promoter. The highly concentrated version and modified T7 RNA Poly-merase HC is ideally suited for set-up and optimization of high yield in vitro transcription reactions. Proline 266 has been replaced by leucine (P266L) which has been associated with: decreased abortive transcription. increased 5' homogeneity of transcripts synthesized from A-initiating phi2.5 promoter; increased 5' incorporation efficiency of GTP analogs The provided T7 IVT Set-up Buffer, MgCl 2and DTT solutions enable testing of different reaction conditions such as nucleotide/MgCl 2 concentrations. Recommendation for assay set-up with minimal optimization: HighYield T7 P&L RNA Synthesis Kit (#orb653830).

Images & Validation

• Application Notes:
  Activity: 1 µl T7 P&L RNA Polymerase HC generates >/= 30 µg RNA in 10 min at 37°. This is comparable to 1000 U of NEB's T7 RNA Polymerase (High Concentration), # M0460T (activity assay conditions 1x T7 IVT Set-up Buffer supplemented with 10 mM DTT, 12 mM MgCl 2, 3 mM NTPs, 1 µl T7 RNA Polymerase HC and 1 µg DNA template (1.4 kpb RNA transcript) in 20 µl. RNA yield was determined with a fluorescence microplate assay.).

Key Properties

• Purity: ≥ 95% (SDS-PAGE)

Storage & Handling

• Storage: store at -20 °C
• Form/Appearance: liquid
• Concentration: 2.0-2.2 µg/µl (A280, ɛ = 140 L mmol-1cm-1 in T7 RNA Polymerase HC Storage Buffer)
• Expiration Date: 12 months from date of receipt.
• Hazard Information: Non-Toxic
• Disclaimer: For research use only

Alternative Names

Highly concentrated and modified T7 RNA Polymerase for set-up of high yield in vitro transcription reactions