US Patent:
20190112655, Apr 18, 2019
Inventors:
- South San Francisco CA, US
Adam R. Sciambi - South San Francisco CA, US
Maurizio Pellegrino - South San Francisco CA, US
Assignee:
Mission Bio, Inc. - South San Francisco CA
International Classification:
C12Q 1/6874
Abstract:
The disclosed embodiments relate to method, apparatus and system for high throughput single-cell DNA sequencing with droplet microfluidic. In an exemplary embodiment, a microfluidic apparatus is used to provide a rapid and cost-effective targeted genomic sequencing of thousands of cells in parallel. The targeted sequencing can be directed for residual disease detection. In one embodiment, the disclosure provides a method to detect one or more mutations in tumor cells, the method comprising: encapsulating at least one cell and a lysis reagent in a carrier fluid to form a droplet, wherein the cell originates from a tumor and the cell comprises a genomic DNA; lysing the cell to release the genomic DNA and thereby form a droplet containing the genomic DNA; introducing a cell identifier and one or more primers specific to a plurality of regions of the genomic DNA; and thermocycling the droplet to amplify the genomic DNA and to incorporate cell identifiers into the genomic DNA to produce a plurality of amplified DNA with identified loci; wherein once the cell identifier is incorporated into the amplified DNA, the identified loci are sequenced and at least one DNA mutation is identified for the tumor cells.