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US 12,577,563 B2Granted
Methods for multiplexing recombinase polymerase amplification
AssigneeAbbott Diagnostics Scarborough, Inc.
InventorsOlaf Piepenburg, Colin H. Williams, Niall A. Armes
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Abstract
This disclosure provides for methods and reagents for rapid multiplex RPA reactions and improved methods for detection of multiplex RPA reaction products. In addition, the disclosure provides new methods for eliminating carryover contamination between RPA processes.
Claims18
A method comprising:
- (a) providing a liquid sample comprising a target nucleic acid;
- (b) combining the liquid sample with a recombinase polymerase amplification (RPA) reagent composition to provide a reaction mixture, wherein the RPA reagent composition comprises:
- (i) a recombinase;
- (ii) a polymerase;
- (iii) a first nucleic acid primer;
- (iv) a second nucleic acid primer;
- (v) a third extension blocked primer comprising an elongation blocking group at a 3′ end, a D-Spacer, and an internal residue non-complementary to the target nucleic acid;
- and
- (vi) a nuclease;
- and
- (c) performing RPA on the reaction mixture to produce an amplicon indicative of the presence of the target nucleic acid.
8 more elements
Dependencies are inferred from the claim text.