HighYield T7 ARCA mRNA Synthesis Kit (Ψ-UTP), S pack

HighYield T7 ARCA mRNA Synthesis Kit (Ψ-UTP) is designed to produce large amounts of ARCA-capped and pseudouridine-modified (m)RNA via in vitro transcription with T7 RNA polymerase. The resulting 5’-capped and internally modified (m)RNA can subsequently be used for microinjection, transfection or in vitro translation experiments. Anti-reverse cap analog (ARCA, m7,3′-OGpppG)-capped (m)RNA possesses a significantly higher translation efficiency compared to traditional m7GpppG-capped (m)RNA. This is due to 3′-O-methylation of the m7 guanine moiety that allows ARCA incorporation in the correct (“anti-reverse”) orientation only resulting in a 100 % translatable capped (m)RNA population. Pseudouridine modifications have been shown to increase (m)RNA stability and to reduce immunogenicity. The kit contains sufficient reagents for 15 reactions (S-Pack) or 50 reactions (L-Pack) à 20 μl (6 mM ARCA, 1.5 mM GTP, 7.5 mM Pseudo-UTP, 7.5 mM ATP, 7.5 mM CTP). An individual optimization of ARCA and Pseudo-UTP concentration can easily be achieved with the single nucleotide format. A 20 µl reaction yields about 30-50 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)).