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

HighYield T7 mRNA Synthesis Kit (m5CTP/Ψ-UTP) is designed to produce large amounts of 5-methylcytidine and pseudouridine-modified (m) RNA via in vitro transcription with T7 RNA polymerase. The resulting internally modified (m) RNA can subsequently be used for microinjection, transfection or in vitro translation experiments. 5-Methylcytidine and pseudouridine 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 labeling reactions à 20 μl (7.5 mM GTP, 7.5 mM 5-Methyl-CTP, 7.5 mM Pseudo-UTP, 7.5 mM ATP) . An individual optimization of 5-methyl-CTP and Pseudo-UTP concentration as well cap analoga incorporation 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) ) .