3 replies to this topic

[xyne]

Hi there,

i need to synthesize a circular RNA and i have the sequence of the circular RNA. Does anyone know how to do it?

thanks.

[phage434]

T4 RNA ligase (different from T4 DNA ligase) will ligate RNA molecules with a 5' phosphate.  Presumably using this enzyme at low RNA concentration to favor circularization over concatamers would work.  How will you tell?  Why do you want to do this?

[xyne]

Thanks.

Actually i just want to transfect the circular RNA with specific sequence inside a cell and study the effect.
Can i know how to extract the circular RNA that i want effectively after the ligation?

Thanks.

[phage434]

There are probably 5' or 3' RNA exonucleases that would selectively degrade the single stranded RNA species.  I don't know how you would quantify or optimize this reaction to make certain the reaction went to completion.