CPC G01N 33/53 (2013.01) [C12N 15/1075 (2013.01); C12Q 1/6804 (2013.01); C12Q 1/6806 (2013.01); C12Q 1/6874 (2013.01); C40B 50/08 (2013.01); G01N 33/532 (2013.01); G01N 33/5436 (2013.01); G01N 33/58 (2013.01); C12Q 2521/10 (2013.01); C12Q 2563/179 (2013.01); C12Q 2565/30 (2013.01); G01N 2458/10 (2013.01)] | 18 Claims |
1. A method for analyzing proteins of a cell, the method comprising:
segregating a single cell and barcoded binders, each barcoded binder comprising a binder linked to an oligonucleotide comprising a binder-identifying barcode, into a first droplet in an immiscible carrier fluid within a microfluidic channel, wherein the binder-identifying barcodes are sticky-ended barcodes made by a process that includes rounds of successive addition of sticky-ended partial barcodes all performed by microfluidic droplet merger;
binding the barcoded binders to proteins of the single cell;
providing a second droplet from a universal barcode droplet library into the microfluidic channel, where the second droplet includes a plurality of copies of a droplet-identifying barcode;
merging the second droplet into the first droplet;
attaching the copies of the droplet-identifying barcode to the binder-identifying barcodes of the barcoded binders bound to the proteins of the single cell to form composite barcodes;
sequencing the composite barcodes; and
identifying the binders from the sequences of the composite barcodes, thereby identifying the proteins of the single cell.
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