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US 12,692,536 B2Granted
Systems and methods for processing optically tagged beads
Assignee10x Genomics, Inc.
InventorsRajiv Bharadwaj, Zachary Bent
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Abstract
The present disclosure provides methods and systems for generating and processing optically tagged beads. A nucleic acid probe comprising a fluorescent tag may be hybridized to a bead to generate an optically tagged bead. The nucleic acid probe hybridized to the optically tagged bead may be detectable by one or more signals and may be substantially unreactive to one or more chemical reactions performed with, or in the presence of, the optically tagged bead.
Claims17
A method, comprising:
- (a) hybridizing a bead and a probe to generate an optically-tagged bead, wherein the optically-tagged bead comprises a binding sequence,
- wherein the probe hybridizes to the binding sequence,
- wherein the probe comprises:
- (i) a fluorescent tag;
- and (ii) at a 3′ end of the probe, a moiety selected from the group consisting of a fluorophore, a carbon spacer, a biotin molecule, an inverted terminal base, a sequence that destabilizes hybridization between the bead and the probe, and an unlocked nucleic acid, and
- wherein the moiety renders the probe hybridized to the bead at least 60% less reactive in a base incorporation reaction as compared to an equivalent nucleic acid probe lacking the moiety;
- and
- (b) detecting one or more signals indicative of the fluorescent tag from the optically-tagged bead,
- wherein the optically-tagged bead comprises:
- i) a molecule comprising the binding sequence hybridized to the probe, and ii) a nucleic acid barcode molecule, wherein the molecule is different from the nucleic acid barcode molecule.
7 more elements
Dependencies are inferred from the claim text.