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Method of sequence determination using sequence tags

  • US 9,506,119 B2
  • Filed: 03/15/2013
  • Issued: 11/29/2016
  • Est. Priority Date: 11/07/2008
  • Status: Active Grant
First Claim
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1. A method of determining clonotypes of an immune repertoire, the method comprising the steps:

  • (a) attaching sequence tags to molecules of recombined nucleic acids of I-cell receptor genes or immunoglobulin genes of T-cells and/or B-cells from a sample comprising cells and/or B-cells to form tag-molecule conjugates, wherein substantially every molecule of the tag-molecule conjugates has a unique sequence tag;

    (b) amplifying the tag-molecule conjugates;

    (c) sequencing by synthesis the tag-molecule conjugates to produce sequence reads comprising at least 1000 clonotypes per run, wherein each sequence read comprises a sequence tag portion and a clonotype portion; and

    (d) aligning like sequence tag portions of the sequence reads to determine a clonotype sequence from corresponding clonotype portions of the aligned sequence reads.

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