Sepharose-supported DNA as template for RNA synthesis

Abstract

A 96-bp DNA fragment containing the bacteriophage SP6 promoter was covalently coupled to EAH-Sepharose 4B in an orientation-specific manner. Here we show that the immobilized DNA was able to serve as template in in vitro transcription of RNA. This solid-phase approach allowed easy separation of the RNA products by centrifugation and made digestion of the template DNA by DNase superfluous. The Sepharose-bound DNA template was active in at least four consecutive cycles of in vitro transcription whose results were highly reproducible. The system may form a useful basis for the development of other molecular biology reactions on solid supports. Copyright (C) 1998 Elsevier Science B.V.