Novel enzymatic methods to generate high yields of sequence specific rna oligonucleotides with extreme precision

A method of synthesizing a product RNA, the method comprising

• preparing a reaction mixture comprising a functional template DNA for the product RNA, and an RNA polymerase under conditions for RNA synthesis, wherein the RNA polymerase is covalently or noncovalently linked to a nontemplate DNA that is 10 nucleotides or longer in length, and wherein the nontemplate DNA is complementary to a minimum of 10 nucleotides of the template DNA upstream of the transcription start site, or wherein the nontemplate DNA is complementary to all or part of the entire template strand DNA in the coding region, wherein the nontemplate DNA and the RNA polymerase are immobilized to a solid support, and wherein the reaction mixture and the solid support are contained in a reaction chamber of a fluidic chip, the fluidic chip comprising fluidic paths for reagent delivery and product RNA removal, and
• synthesizing the product RNA in the reaction mixture and continuously removing the product RNA from the reaction chamber while flowing fresh RNA synthesis reagents into the reaction chamber,
• wherein the RNA polymerase comprises an avidin-binding peptide, the nontemplate DNA comprises a 3′ or 5′ biotin label respectively, and the RNA polymerase is noncovalently linked to the nontemplate DNA via a bead support that binds both the avidin-binding peptide and the biotin.