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US 2023/0416809 A1Published application
Spatial detection of biomolecule interactions
AssigneeSingular Genomics Systems, Inc.
InventorChristian BERRIOS
Life of this patent
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Abstract
Disclosed herein, inter alia, are compositions and methods for spatial detection of biomolecular interactions.
Claims20
A method of forming an oligonucleotide comprising two barcode sequences, said method comprising:
- a) contacting a first biomolecule with a first proximity probe, wherein the first proximity probe comprises a first oligonucleotide comprising, from 5′ to 3′, a first primer binding sequence, a first barcode sequence, and a first probe sequence;
- b) contacting a second biomolecule with a second proximity probe, wherein the second proximity probe comprises a second oligonucleotide comprising, from 5′ to 3′, a second primer binding sequence, a second barcode sequence, and a second probe sequence;
- c) hybridizing the first probe sequence of said first oligonucleotide to the second probe sequence of said second oligonucleotide and extending the first probe sequence with a polymerase to form a first extended oligonucleotide comprising, from 5′ to 3′, the first primer binding sequence, the first barcode sequence, the first probe sequence, a complement of the second barcode sequence, and a complement of the second primer binding sequence.
The method of claim 16, wherein said cleavable linker comprises a polynucleotide or a polypeptide sequence.
A composition comprising:
- i) a biomolecule bound to a proximity probe, wherein the proximity probe comprises an extended probe oligonucleotide comprising, from 5′ to 3′, a first primer binding sequence, a first barcode sequence, a first probe sequence, a complement of a second barcode sequence, and a complement of a second primer binding sequence;
- and
- ii) an oligonucleotide primer hybridized to the extended probe oligonucleotide, wherein the oligonucleotide primer comprises, from 5′ to 3′, a first sequence complementary to the first primer binding sequence and a second sequence complementary to the complement of the second primer binding sequence.
A composition comprising:
- i) a biomolecule bound by a proximity probe, wherein the proximity probe comprises an extended probe oligonucleotide comprising, from 5′ to 3′, a first primer binding sequence, a first barcode sequence, a first probe sequence, a complement of a second barcode sequence, a complement of a third probe sequence, a complement of a third barcode sequence, a complement of a fifth probe sequence, an internal cleavable site, and a complement of a second primer binding sequence;
- and
- ii) an oligonucleotide primer hybridized to the extended probe oligonucleotide, wherein the oligonucleotide primer comprises, from 5′ to 3′, a first sequence complementary to the first primer binding sequence and a second sequence complementary to the complement of the second primer binding sequence.
Dependencies are inferred from the claim text.