In vitro transcription of mRNA with the NEB HiScribe™ SP6 Quick RNA Synthesis Kit and subsequent enzymatic capping v1

This a protocol for in vitro transcription (IVT) of mRNA with SP6 RNA polymerase (with or without template-based polyA-tail) followed by enzymatic capping. A single reaction should produce between 60 and 160 µg mRNA. The protocol was used to produce mRNA of different fluorescent proteins for electroporation of Aiptasia (Exaiptasia diaphana) zygotes. There, a simple cap0 5′ structure and a polyA tail of 50 nt proved most efficient for expression in vivo, this might be different in other organisms.