HighYield T7 mRNA Synthesis Kit (m5CTP), L pack

HighYield T7 mRNA Synthesis Kit (m5CTP) is designed to produce large amounts of 5-methylcytidine-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. 5-Methylcytidine modifications have been shown to increase (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 UTP, 7.5 mM ATP, 7.5 mM 5-Methyl-CTP). An individual optimization of 5-Methyl-CTP concentration can easily be achieved with the single nucleotide format. A 20 µl reaction yields about 100-130 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)).