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US 11,891,666 B2Granted
Methods and systems for detecting biological components
AssigneeThe Regents of the University of California
InventorsAdam R. Abate, Dennis Jay Eastburn, Adam R. Sciambi
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Abstract
Methods for the detection of components from biological samples are provided. In certain aspects, the methods may be used to detect and/or quantify specific components in a biological sample, such as tumor cells (e.g., circulating tumor cells). Systems and devices for practicing the subject methods are also provided.
Claims10
A method for genotyping a cell, the method comprising:
- (a) encapsulating lysis reagents and the cell containing a target polynucleotide in a droplet, the lysis reagents comprising an enzyme having protease activity, wherein the lysate droplet is surrounded by an immiscible carrier fluid;
- (b) incubating the droplet containing the cell and the lysis reagents comprising the enzyme having protease activity for a period of time sufficient to lyse the cell and form a cell lysate comprising the target polynucleotide and for the protease to digest inhibitory proteins;
- (c) inactivating the enzyme having protease activity;
- (d) introducing amplification reagents to form a droplet containing the cell lysate and the amplification reagents;
- and
- (e) amplifying the target polynucleotide or a complement of the target polynucleotide within the droplet containing the cell lysate and the amplification reagents, wherein the method does not comprise a step of selectively removing reagents from the droplet containing the cell lysate and amplification reagents prior to step (e);
- and
- (f) sequencing the amplified target polynucleotide or amplified complement of the target polynucleotide, thereby detecting a presence or absence of one or more changed nucleotide bases in the target polynucleotide.
5 more elements
Dependencies are inferred from the claim text.