CPC G01N 33/6878 (2013.01) [B01J 19/0046 (2013.01); C12Q 1/6816 (2013.01); C12Q 1/6869 (2013.01); C40B 20/04 (2013.01); C40B 70/00 (2013.01)] | 9 Claims |
1. A method for generating a plurality of mixture partitions, the method comprising:
(i) providing a plurality of fixed and permeabilized single cells;
(ii) incubating the plurality of single cells with a library of at least 10 structurally distinct antibodies that have a specific binding affinity for target epitopes of cellular proteins, wherein each of the at least 10 antibodies is conjugated to a different epitope-specific oligonucleotide comprising a barcode sequence that identifies the target epitope, a unique molecular identifier (UMI), a first primer binding sequence, and a second primer binding sequence, wherein the incubating results in binding of the antibodies to their corresponding epitopes, if present, to form antibody-epitope complexes;
(iii) washing away unbound antibodies;
(iv) partitioning, after the incubating and washing steps, the plurality of fixed and permeabilized single cells into a plurality of mixture partitions; and
(v) providing a plurality of partition-specific oligonucleotides to each mixture partition, wherein each partition-specific oligonucleotide comprises a different barcode sequence and a primer binding sequence that is complementary to the first primer binding sequence of the epitope-specific oligonucleotides,
wherein each partition-specific oligonucleotide is covalently linked to a bead.
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