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US 11,299,766 B2Granted
Multiplex amplification detection assay
AssigneeExact Sciences Corporation
InventorsGraham P. Lidgard, Hatim Allawi
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Abstract
Provided herein is technology relating to the amplification-based detection of bisulfite-treated DNAs and particularly, but not exclusively, to methods and compositions for multiplex amplification of low-level sample DNA prior to further characterization of the sample DNA. The technology further provides methods for isolating DNA from blood or blood product samples, e.g., plasma samples.
Claims20
A method of analyzing a sample for multiple target nucleic acids in a PCR-flap assay, comprising:
- a) providing a sample comprising DNA suspected of containing one or more of a plurality of different target regions;
- b) treating the sample in an amplification reaction mixture comprising a PCR-flap assay buffer comprising 6 to 10 mM Mg′ under conditions wherein the different target regions, if present in the sample, are amplified to form a pre-amplified mixture;
- c) partitioning the pre-amplified mixture into a plurality of different PCR-flap assay reaction mixtures comprising the PCR-flap assay buffer;
- and
- d) conducting a plurality of PCR-flap assays with the PCR-flap assay reaction mixtures, wherein the different target regions, if present in the sample at step a), are amplified and detected in the PCR-flap assay reaction mixtures.
2 more elements
Dependencies are inferred from the claim text.