How can we sythesize 6 kb mRNA in vitro?

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Junjie Shao

lots of T7/SP6 transcription kit works.

invitrogen, neb, etc.

you would need to generate a plasmid with T7 promoter and your GOI.

Wei Li

But, 6 kb is too long. There would be lots of short and incomplete RNA. Production of complete 6 kb RNA is quite low. A friend sythesize 3.5 kb SpCas9 mRNA by T7 kit produce lots of short RNA.

Did you try T7 in vitro synthesize 6kb RNA?

Michele Gabriele

Wei Li I even managed to to in vitro transcription with of an mRNA about 10kb long, with similar kits that Junjie Shao was mentioning. Have a look here for the methods Article Efficient Generation of Human iPSCs by a Synthetic Self-Repl...

you should change the gel and the buffer for the gel and use specific products for conservation