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US 2026/0028619 A1Published application
Methods, compositions, and kits for single cell analysis by tagmentation
Assignee10x Genomics, Inc.
InventorsPaulius Mielinis, Marlon Stoeckius, David Sukovich, Marco Serra, Lauren Gutgesell
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Abstract
Provided herein are methods, systems, and kits for detecting analytes of interest from biological samples using tagmentation. The methods include tagmenting DNA/RNA hybrids that include an mRNA molecule and a barcode nucleic acid molecule to generate a plurality of tagmented fragments, and separating a first tagmented fragment including an end portion of the mRNA and a barcode from other tagmented fragments.
Claims28
A method for processing an mRNA molecule, the method comprising:
- (a) in a partition, hybridizing the mRNA molecule to a barcode nucleic acid molecule attached to a solid support, wherein the barcode nucleic acid molecule comprises:
- (i) a barcode sequence and (ii) a domain that hybridizes to a 3′ sequence of the mRNA molecule;
- (b) extending the barcode nucleic acid molecule, thereby generating a barcoded nucleic acid molecule annealed to the mRNA molecule, wherein the barcoded nucleic acid molecule comprises a sequence complementary to the mRNA molecule;
- (c) tagmenting the barcoded nucleic acid molecule annealed to the mRNA molecule, thereby generating a plurality of tagmented fragments comprising a first tagmented fragment, wherein the first tagmented fragment comprises a first strand comprising an mRNA fragment comprising 3′ sequence of the mRNA molecule covalently attached to the transposon end sequence, and a second strand comprising the barcode nucleic acid molecule and a portion of the sequence complementary to the mRNA molecule;
- and
- (d) separating the first tagmented fragment from other tagmented fragments of the plurality of tagmented fragments.
3 more elements
Dependencies are inferred from the claim text.