US 11,747,327 B2
Compositions and methods for molecular labeling
Michael L. Samuels, Windham, NH (US); Jeffrey Charles Olson, Chelmsford, MA (US); Andrew Watson, Bedford, MA (US); Keith Brown, Carlsbad, CA (US); and Darren R. Link, Lexington, MA (US)
Assigned to BIO-RAD LABORATORIES, INC., Hercules, CA (US)
Filed by Bio-Rad Laboratories, Inc., Hercules, CA (US)
Filed on Jul. 14, 2020, as Appl. No. 16/928,924.
Application 16/928,924 is a continuation of application No. 16/795,467, filed on Feb. 19, 2020.
Application 16/795,467 is a continuation of application No. 16/400,858, filed on May 1, 2019, granted, now 11,168,353.
Application 16/400,858 is a continuation of application No. 14/995,744, filed on Jan. 14, 2016, granted, now 10,584,332.
Application 14/995,744 is a continuation of application No. 14/874,553, filed on Oct. 5, 2015.
Application 14/874,553 is a continuation of application No. 13/398,677, filed on Feb. 16, 2012, granted, now 9,150,852, issued on Oct. 6, 2015.
Claims priority of provisional application 61/476,714, filed on Apr. 18, 2011.
Claims priority of provisional application 61/444,612, filed on Feb. 18, 2011.
Prior Publication US 2020/0399635 A1, Dec. 24, 2020
Int. Cl. G01N 33/53 (2006.01); G01N 33/532 (2006.01); C40B 50/08 (2006.01); C12Q 1/6874 (2018.01); G01N 33/543 (2006.01); G01N 33/58 (2006.01); C12Q 1/6804 (2018.01); C12Q 1/6806 (2018.01); C12N 15/10 (2006.01)
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
OG exemplary drawing
 
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.