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Strategy for the production of RNA from immobilized templates

  • US 5,700,667 A
  • Filed: 05/18/1995
  • Issued: 12/23/1997
  • Est. Priority Date: 09/20/1993
  • Status: Expired due to Fees
First Claim
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1. A method of producing RNA enzymatically in a bioreactor comprising the steps of:

  • (a) immobilizing a DNA template, comprising a noncoding strand and a coding strand comprising a double-stranded promoter sequence and a double-stranded spacer sequence upstream of said promoter sequence, to a solid support through a linkage comprising a single-stranded overhang extending from the 5'"'"'-end of said noncoding strand and such that said coding strand remains dissociable from said noncoding strand;

    (b) contacting said immobilized DNA template of step (a) with a transcription reaction mixture comprising a buffer, nucleoside triphosphates and an RNA polymerase such that RNA having a sequence complementary to said coding strand is produced;

    (c) removing said RNA produced in step (b); and

    ,(d) repeating steps (b) and (c).

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