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US 12,735,748 B2Granted
Methods for simultaneous amplification of target loci
AssigneeNatera, Inc.
InventorsMatthew Rabinowitz, Matthew Micah Hill, Bernhard Zimmermann, Johan Baner, George Gemelos, Milena Banjevic, Allison Ryan, Styrmir Sigurjonsson, Zachary Demko
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Abstract
The invention provides methods for simultaneously amplifying multiple nucleic acid regions of interest in one reaction volume as well as methods for selecting a library of primers for use in such amplification methods. The invention also provides library of primers with desirable characteristics, such as minimal formation of amplified primer dimers or other non-target amplicons.
Claims9
A method for amplifying and sequencing DNA, comprising:
- amplifying isolated cell-free DNA or DNA derived therefrom one or more times to generate amplification products, wherein the isolated cell-free DNA is isolated from a blood sample collected from a subject who is not a pregnant woman, wherein one of the amplification steps comprises targeted amplification of a plurality of single nucleotide polymorphism or variant loci in a single reaction volume;
- tagging the amplification products with one or more universal tail adaptors to generate tagged products, and amplifying the tagged products using adaptor-specific primers to generate final amplification products, wherein the final amplification products comprise a barcode and one or more sequencing tags, wherein tagging the amplification products comprises appending the one or more universal tail adaptors to the amplification products;
- and
- sequencing the plurality of single nucleotide polymorphism or variant loci on the cell free DNA by conducting massively parallel sequencing on the final amplification products, wherein the plurality of single nucleotide polymorphism or variant loci comprises 50-2,000 loci associated with cancer.
1 more element
Dependencies are inferred from the claim text.