HighYield T7 ARCA mRNA Synthesis Kit (m5CTP/Ψ-UTP)

HighYield T7 ARCA mRNA Synthesis Kit (m5CTP/Ψ-UTP) is designed to produce large amounts of ARCA-capped, 5-methylcytidine 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 m7guanine moiety that allows ARCA incorporation in the cor-rect ("anti-reverse") orientation only resulting in a 100% translatable capped (m) RNA population. 5-Methylcytidine and pseudouridine modifications have been shown to increase (m) RNA stability and to reduce immunogenicity. The kit contains sufficient reagents for 15 labeling reactions à 20 μl (6 mM ARCA, 1.5 mM GTP, 7.5 mM 5-Methyl-CTP, 7.5 mM Pseudo-UTP, 7.5 mM ATP) . An individual optimization of ARCA, 5-methyl-CTP 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 incu-bation (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) ) .