In vitro synthesis of RNA (> 20 nt up to several thousand nt) is catalyzed by bacteriophage RNA polymerases using linear DNA as a template (in vitro transcription). T7 RNA polymerase is the most efficient and widely used RNA polymerase. A modified version (T7 P&L RNA polymerase) with proline 266 replaced by leucine (P266L) has been associated with decreased abortive transcription[1], increased 5' homogeneity of transcripts synthesized from A-initiating phi2.5 promoter[2], increased 5' incorporation efficiency of GTP analogs[3].
140 - 160 µg RNA are synthesized after 30 min incubation with our HighYield formulation (1 μg T7 control template, 1.4 kb RNA transcript).
[1] Guillerez et al. (2005) A mutation in T7 RNA polymerase that facilitates promoter clearance. Natl. Acad. Sci. U.S.A102:5958.
[2] Salvail-Lacoste et al. (2018) Affinity purification of T7 RNA transcripts with homogeneous ends using ARiBo and CRISPR tags. RNA19:1003.
[3] Lyon et al. (2018) A T7 RNA Polymerase Mutant Enhances the Yield of 5'-Thienoguanosine-Initiated RNAs. ChemBioChem19:142.
