Patent ID: 11891666
Assignee: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Field: Chemical engineering (Chemistry)
Classification: CPC C  B | IPC B  C

Claim 0:
1. 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.