10X Genomics, Inc.

Paul Eugene Lund

Katherine Pfeiffer

Andrew Kohlway

Andrew Hill

Shankar Shastry

Javelin C Chi

Disclosed herein is a method of preventing the amplification of artifactual antisense products when sequencing a target polynucleotide. Incorporation of modified nucleotides into handle polynucleotide sequences can allow for the selective degradation of artifact antisense products generated by mis-priming, prior to amplification, thus preventing them from appearing in the amplification library. Selective degradation can be accomplished using one or more agents configured to specifically cleave the handle polynucleotide at the modified nucleotide. Amplification can be performed using a primer that is configured to hybridize to the handle sequence and a second primer that is configured to hybridize to a complement of a target nucleotide.

Methods and systems for reducing artifactual antisense products

Provided herein are systems and methods for processing biomolecules (e.g., nucleic acid molecules, proteins) from a sample. A method for processing biomolecules may comprise hybridizing a probe molecule to a target region of a nucleic acid molecule (e.g., a ribonucleic acid (RNA) molecule) and barcoding the probe-nucleic acid molecule complex or derivatives thereof. Such a method can comprise performing a nucleic acid reaction, e.g., extension, denaturation, and amplification. A method for processing a sample may comprise hybridizing probes to (i) target regions of a nucleic acid molecule (e.g., RNA molecule) and (ii) a reporter oligonucleotide of a feature binding group, and barcoding the probe-associated molecules. One or more processes of the methods described herein may be performed within a partition, such as a droplet or well.

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San Leandro, CA, United States of America

Paul Eugene Lund