HighYield T7 ARCA mRNA Synthesis Kit (m1ATP)

HighYield T7 ARCA mRNA Synthesis Kit (N1-Methyl-ATP) is designed to produce large amounts of ARCA-capped and N1-methyladenosine-modified (m) RNA via in vitro transcription with T7 RNA polymerase. The resulting 5'-capped and internally modified (m) RNA can subse-quently 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. N1-methyladenosine 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 UTP, 7.5 mM CTP, 3.75 mM ATP, 3.75 mM N1-Methyl-ATP) . An individual optimization of ARCA and N1-Methyl-ATP concentration can easily be achieved with the single nucleotide format. A 20 μl reaction yields about 10-30 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) ) .