HighYield T7 mRNA Synthesis Kit (me1Ψ-UTP)

HighYield T7 mRNA Synthesis Kit (me1Ψ-UTP) is designed to produce large amounts of N1-Methylpseudouridine-modified (m) RNA via in vitrotranscription with T7 RNA polymerase. The resulting internally modified (m) RNA can subsequently be used for microinjection, transfection or in vitro translation experiments. N1-Methylpseudouridine modifications have been shown to in-crease (m) RNA stability and to reduce immunogenicity. A 5'-cap structure further increases translation efficiency. 5'-capping can be achieved by a) co-transcriptional capping with cap analoga (e.g. ARCA) or b) post-transcriptional capping with capping enzymes. The kit contains sufficient reagents for 15 reactions (S-Pack) or 50 reactions (L-Pack) à 20 μl (7.5 mM GTP, 7.5 mM N1-Methylpseudo-UTP, 7.5 mM ATP, 7.5 mM CTP) . An individual optimization of N1-Methylpseudo-UTP concentration can easily be achieved with the single nucleotide format. A 20 μl reaction yields about 100-130 μg RNA after 30 min in-cubation (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) ) .