Hello everyone,
I am working on the production of IVT mRNA. After transcription, the remaining DNA templates are digested by DNase I. I have tested transcription with uncut DNA (mostly supercoiled) and with linearised DNA as it is poorly described in literature, how the efficiency of transcription is reduced by using supercoiled DNA. What I ended up finding upon viewing my transcription products on agarose gels, was incomplete digestion of the DNA when it was in its supercoiled form. So I am now taking a closer look at that. Can anyone advise how the form of DNA impacts the efficiency of DNase I digestions?