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US 2014/0155274 A1Published application
Single Cell Nucleic Acid Detection and Analysis
AssigneePresident and Fellows of Harvard College
InventorsXiaoliang Sunney Xie, Katsuyuki Shiroguchi, Peter A. Sims, Tony Z Jia
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Abstract
Methods and compositions for digital profiling of nucleic acid sequences present in a sample are provided.
Claims24
A method of determining copy number of a nucleic acid molecule in a sample including a plurality of nucleic acid molecules comprising
- attaching a unique barcode sequence to substantially each of the plurality of nucleic acid molecules in the sample to produce a plurality of barcoded nucleic acid molecules,
- amplifying the plurality of barcoded nucleic acid molecules in the sample to produce amplicons of the plurality of barcoded nucleic acid molecules,
- sequencing each amplicon to identify an associated nucleic acid sequence and an associated barcode sequence,
- selecting a first target nucleic acid sequence and determining the number of unique associated barcode sequences for the first target nucleic acid sequence, wherein the number of unique associated barcode sequences is the copy number of the first target nucleic acid sequence.
1 more element
A method of counting nucleic acid molecules in a sample including a plurality of nucleic acid molecules comprising
- attaching a unique barcode sequence to substantially each of the plurality of nucleic acid molecules in the sample to produce a plurality of barcoded nucleic acid molecules,
- amplifying the plurality of barcoded nucleic acid molecules in the sample to produce amplicons of the plurality of barcoded nucleic acid molecules,
- sequencing each amplicon to identify an associated barcode sequence, and
- counting the number of unique associated barcode sequences as a measure of the number of nucleic acid molecules in the sample.
1 more element
A method of determining copy numbers of nucleic acid molecules in a sample comprising
- attaching a unique barcode sequence to substantially each of the nucleic acid molecules in the sample to produce a plurality of barcoded nucleic acid molecules,
- amplifying the plurality of barcoded nucleic acid molecules in the sample to produce amplicons of the plurality of barcoded nucleic acid molecules,
- massively parallel sequencing the amplicons of the plurality of barcoded nucleic acid molecules to identify for each amplicon an associated nucleic acid sequence and an associated barcode sequence, and
- determining the number of unique associated barcode sequences for each nucleic acid sequence in the sample.
1 more element
Dependencies are inferred from the claim text.