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US 11,898,205 B2Granted
Increasing capture efficiency of spatial assays
Assignee10x Genomics, Inc.
InventorFelice Alessio Bava
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Abstract
This disclosure relates to methods for increasing capture efficiency of a spatial array using rolling circle amplification of a padlock probe that hybridizes to a capture probe. Also provided are methods for using such spatial arrays to detect a biological analyte in a biological sample.
Claims16
A method for preparing a spatial array, the method comprising:
- (a) contacting a substrate comprising a plurality of capture probes with a padlock probe, wherein:
- (i) a capture probe of the plurality of capture probes comprises a first docking sequence, a spatial barcode, a capture domain, and a second docking sequence;
- and
- (ii) the padlock probe comprises:
- a first docking padlock sequence at its 3′ end that is complementary to the first docking sequence of the capture probe, and
- a second docking padlock sequence at its 5′ end that is complementary to the second docking sequence of the capture probe;
- (b) hybridizing the first docking sequence to the first docking padlock sequence and hybridizing the second docking sequence to the second docking padlock sequence;
- (c) ligating the padlock probe, thereby generating a ligated padlock probe that comprises the capture domain or a complement thereof;
- and
- (d) amplifying the ligated padlock probe, thereby generating an amplified padlock probe sequence with multiple copies of the capture domain.
7 more elements
Dependencies are inferred from the claim text.