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

Biorbyt

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

16

Research Use Only

HighYield T7 ARCA mRNA Synthesis Kit (m5CTP/psi-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 a 20 microl (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 microl reaction yields about 30-50 microg RNA after 30 min incu-bation (1 microg T7 control template, 1.4 kb RNA transcript). Yields may however vary depending on the template (promotor design, sequence length, secondary structure formation)).

-20°C

12352200