cDNA SPIKE-IN CONTROL FOR SINGLE CELL ANALYSIS
AssigneeBecton, Dickinson and Company
InventorsTracy Campbell, Hye-Won Song
Life of this patent
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Abstract
Disclosed herein include systems, methods, compositions, and kits for determining the presence of a single cell mRNA sequencing assay workflow failure, including determining a failure in barcoding copies of a nucleic acid target and determining a failure in sequencing library generation. There are also provided, in some embodiments, compositions, methods, and systems for determining the sequencing status of sequencing library members (e.g., saturated sequencing or under sequencing). Compositions comprising a predetermined copy number of barcoded control nucleic acids are also provided herein.
Claims20
A method for labeling nucleic acid targets in a sample, comprising:
- barcoding copies of a nucleic acid target with a first plurality of oligonucleotide barcodes to generate a plurality of barcoded nucleic acid molecules each comprising a sequence complementary to at least a portion of the nucleic acid target;
- providing a plurality of one or more barcoded control nucleic acids, wherein the number of copies of each of the one or more barcoded control nucleic acids is predetermined;
- generating a sequencing library comprising a plurality of nucleic acid target library members and a plurality of control nucleic acid library members, wherein generating a sequencing library comprises:
- attaching sequencing adaptors to the plurality of barcoded nucleic acid molecules, or products thereof, to generate the plurality of nucleic acid target library members;
- and
- attaching sequencing adaptors to the plurality of one or more barcoded control nucleic acids, or products thereof, to generate the plurality of control nucleic acid library members;
- and
- obtaining sequencing data comprising a plurality of sequencing reads of one or more nucleic acid target library members and a plurality of sequencing reads of one or more control nucleic acid library members.
A kit comprising:
- a plurality of one or more barcoded control nucleic acids, wherein the number of copies of each of the one or more barcoded control nucleic acids is predetermined, wherein the plurality of one or more barcoded control nucleic acids are associated with a second solid support.
Dependencies are inferred from the claim text.