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US 2018/0094288 A1Published application
Methods for the synthesis of polyadenylic acid
AssigneeIbis Biosciences, Inc.
InventorsMark Eshoo, Curtis Phillipson
Life of this patent
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Abstract
The present invention provides compositions, kits, and methods for synthesizing polyadenylic acid using polynucleotide phosphorylase, adenosine diphosphate, a buffering agent, and a divalent metal cation. In certain embodiments, the adenosine diphosphate is present at a concentration between about 5.0 mM and about 100 mM, and the buffering agent has a pH around 8.0. In some embodiments, the prepared polyadenylic acid compositions are free of detectable nucleic acid.
Claims10
A method of making unlabeled polyadenylic acid, comprising:
- a) combining polynucleotide phosphorylase, unlabled adenosine diphosphate (ADP) molecules, a buffering agent with pH between 7.4 and 8.3, and a divalent metal cation, to generate a mixture, wherein said ADP molecules are present in said mixture at a concentration between 30 mM and 50 mM, wherein each of said ADP molecules consist of the following structure:
- and
- b) incubating said mixture under conditions such that a composition comprising unlabeled polyadenylic acid molecules is generated, wherein said unlabeled polyadenylic acid molecules each consist of a plurality of adenylic acid residues, wherein said incubating is conducted for 100 hours to 150 hours at said pH between 7.4 and 8.3, and wherein said composition is free of detectable contaminating nucleic acid.
Dependencies are inferred from the claim text.