HighYield T7 P&L RNA NMR Kit (5F-UTP)

SKU: orb692280

Description

HighYield T7 P&L RNA NMR Kit (5F-UTP) is designed to produce large amounts of 5-Fluoro-modified RNA via in vitro transcription with T7 RNA polymerase. The resulting internally modified (m)RNA can subsequently be used for structure determination vianuclear magnetic resonance (NMR) spectroscopy. 5F-UTP is incorporated instead of its natural counterpart in a non-perturbing way (intact base pairing properties). 5-Fluoro-modification simultaneously functions as a sensitive NMR reporter group (larger chemical shift dispersion than1H). The modified T7 RNA polymerase (T7 P&L RNA polymerase) with proline 266 replaced by leucine (P266L) has been associated with increased 5' homogeneity of transcripts especially when synthesized from A-initiating phi2.5 promoter. The kit contains sufficient reagents for 15 reactions (S-Pack) or 50 reactions (L-Pack) á 20 µl (7.5 mM GTP, 7.5 mM 5-Fluoro-UTP, 7.5 mM ATP, 7.5 mM CTP). An individual optimization of 5-Fluoro-UTP concentration can easily be achieved with the single nucleotide format. A 20 µl reaction yields about 80 µg RNA after 30 min incubation (1 µg T7 control template, 1.4 kb RNA transcript). Yields may however vary depending on the template (promotor design, sequence length, secondary structure formation).

Storage & Handling

• Storage: store at -20 °C
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

Synthesis of 5-Fluoro-modified RNA