In vitro assay buffer for Cas9
AssigneeAgilent Technologies, Inc.
InventorsJeffrey Carl Braman, Yuchu Grace Hsiung, Katherine Felts
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Abstract
Provided herein is a reaction mixture comprising Cas9 and a non-ionic surfactant, e.g., a polyoxyethylene surfactant. In certain embodiments, the reaction mixture may comprise a Cas9 protein, a guide RNA, a salt, a buffering agent, a nucleic acid target and a non-ionic surfactant. Kits are also provided. In certain embodiments, a kit may comprise: a Cas9 protein; and a concentrated reaction buffer comprising salt, a buffering agent and a non-ionic surfactant.
Claims20
A reaction mixture for in vitro cleavage of a nucleic acid target comprising:
- a) a Cas9 protein;
- b) a guide RNA;
- c) a salt;
- d) a buffering agent;
- e) a nucleic acid target;
- and
- f) a non-ionic surfactant, wherein the non-ionic surfactant is a polyoxyethylene or polyoxypropylene surfactant, a glycosidic surfactant, or a bile salt surfactant;
- wherein components a)-f) are combined in vitro.
A reaction mixture for in vitro cleavage of a nucleic acid target comprising:
- a) a Cas9 protein;
- b) a guide RNA;
- c) a salt;
- d) a buffering agent;
- e) a nucleic acid target;
- and
- f) a non-ionic surfactant, wherein the non-ionic surfactant has an HLB in the range of 12-15;
- wherein components a)-f) are combined in vitro.
Dependencies are inferred from the claim text.