Method of performing droplet-based assays

• Main Authors: Colston, Jr., Billy W, Hindson, Benjamin J, Masquelier, Donald A, Ness, Kevin D
• Format: Patent
• Language: English
• Published: 28.03.2023
• Subjects: AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING; BEER; BIOCHEMISTRY; CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; INDEXING SCHEME ASSOCIATED WITH SUBCLASS B29C, RELATING TO PARTICULARARTICLES; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MIXING, e.g. DISSOLVING, EMULSIFYING, DISPERSING; MUTATION OR GENETIC ENGINEERING; PERFORMING OPERATIONS; PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDEDFOR; SHAPING OR JOINING OF PLASTICS; SPIRITS; TESTING; TRANSPORTING; VINEGAR; WINE; WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE, IN GENERAL

Method of analysis. In the method, a first emulsion and a second emulsion substantially separated from one another by a spacer fluid may be formed. The first emulsion, the spacer fluid, and the second emulsion may be flowed in a channel from a fluid inlet to a fluid outlet of a heating and cooling station having two or more temperature-controlled zones, such that each emulsion is thermally cycled to promote amplification of a nucleic acid target in droplets of the emulsion. Amplification data may be collected from individual droplets of each emulsion downstream of the heating and cooling station. A level of the nucleic acid target present in each emulsion may be determined based on the amplification data collected from the individual droplets of the emulsion.