METHODS FOR PRODUCTION AND QUANTIFICATION OF UNIQUE MOLECULAR IDENTIFIER-LABELED BEADS

• Main Authors: BEHLKE, Mark, HENCK, Steven A, ALLEN, Shawn D, DOBOSY, Joseph, ROSE, Scott D, MANTHEY, Jeffrey A
• Format: Patent
• Language: English
• Published: 18.07.2024
• Subjects: BEER; BIOCHEMISTRY; CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOIDCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PERFORMING OPERATIONS; PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; THEIR RELEVANT APPARATUS; TRANSPORTING; VINEGAR; WINE

Described herein are compositions and methods for the production and quantification of barcoded or unique molecular identifier (UMI)-labeled substrates. In one aspect, the substrate is a bead comprising a template oligonucleotide that is elongated by successive extension reactions to provide a bead with an oligonucleotide comprising a plurality of barcodes and conserved anchor regions. The oligonucleotide may also comprise a poly-T region for the capture and subsequent sequencing of mRNA. Methods are also described for quantifying the amount of template oligonucleotide loaded onto the substrate and the products of the extension reaction after each round and after the final extension.