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

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 via nuclear 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 than 1H). 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).