US 12,454,720 B2
Methods and systems for multiplex analysis
Christopher MacDonald, Carlsbad, CA (US); Aditya Rajagopal, Carlsbad, CA (US); and Dominic Yurk, Carlsbad, CA (US)
Assigned to ChromaCode, Inc., Carlsbad, CA (US)
Appl. No. 17/047,160
Filed by ChromaCode, Inc., Carlsbad, CA (US)
PCT Filed Apr. 16, 2019, PCT No. PCT/US2019/027751
§ 371(c)(1), (2) Date Oct. 13, 2020,
PCT Pub. No. WO2019/204357, PCT Pub. Date Oct. 24, 2019.
Claims priority of provisional application 62/659,025, filed on Apr. 17, 2018.
Claims priority of provisional application 62/659,027, filed on Apr. 17, 2018.
Claims priority of provisional application 62/693,754, filed on Jul. 3, 2018.
Claims priority of provisional application 62/753,782, filed on Oct. 31, 2018.
Claims priority of provisional application 62/804,574, filed on Feb. 12, 2019.
Prior Publication US 2021/0254147 A1, Aug. 19, 2021
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6851 (2018.01); C12Q 1/686 (2018.01)
CPC C12Q 1/6851 (2013.01) [C12Q 1/686 (2013.01)] 17 Claims
OG exemplary drawing
 
1. A method of quantifying a nucleic acid target in a sample, the method comprising:
(a) providing a mixture comprising:
i. a plurality of nucleic acid molecules derived from, and/or corresponding with, said nucleic acid target; and
ii. a plurality of oligonucleotide probes, each of which corresponds to a different region of said nucleic acid target;
(b) partitioning said mixture into a plurality of partitions;
(c) generating a plurality of signals in said plurality of partitions,
wherein said plurality of signals are detectable in one color channel, and
wherein at least one signal of said plurality of signals corresponds with a unique combination of two or more of said plurality of nucleic acid molecules in a partition of said plurality of partitions;
(d) detecting from multiple partitions of said plurality of partitions said plurality of signals in said color channel;
(e) using said plurality of signals detected in step (d) to determine, for said plurality of partitions, a plurality of probabilities that one or more nucleic acid molecules of said plurality of nucleic acid molecules is present in a given partition, thereby generating a plurality of probabilities; and
(f) quantifying said nucleic acid target in said sample based at least on a function of said plurality of probabilities,
wherein said method is performed without determining a number of partitions comprising a nucleic acid sequence corresponding to said nucleic acid target.