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Fast HPLC analytics to characterise IVT reaction –detection of pDNA, mRNA, and IVT components

HPLC with convective chromatography media (e.g.monolith) offers a rapid analytical method to characterise complex mixtures. Transcription reaction used for production of mRNA represents such a mixture, with components varying in size, chemical and physical properties. A new analytical HPLC approach (PATfix) presented here uses CIMacPrimaS to separate IVT components such as triphosphate-nucleotides (NTPs), enzymes, DNA template and RNA in a very short gradient.