×

Multiple tagging of long DNA fragments

  • US 10,557,166 B2
  • Filed: 03/17/2014
  • Issued: 02/11/2020
  • Est. Priority Date: 03/15/2013
  • Status: Active Grant
First Claim
Patent Images

1. A method for preparing genomic DNA for sequence analysis, the method comprising:

  • (a) providing a reaction mixture comprising long fragments of the genomic DNA and a population of beads,wherein the reaction mixture is not compartmentalized into aliquots or nanodrops,wherein the reaction mixture comprises more than a haploid amount of said genomic DNA,wherein each bead comprises at least 1000 oligonucleotides immobilized thereon, said oligonucleotides comprising a tag-containing sequence,wherein the oligonucleotides immobilized on the same individual bead comprise the same tag-containing sequence,wherein each tag-containing sequence comprises a tag sequence,wherein the population of beads comprises, in aggregate, at least 1000 different tag sequences, andwherein the long fragments are from 5 kilobases to 750 kilobases in length;

    (b) producing tagged long fragments by incorporating multiple copies of a tag sequence into at least some of the long fragments, wherein in a plurality of the tagged long fragments said multiple copies are from a single bead;

    (c) producing a plurality of tagged subfragments of the tagged long fragments, wherein a plurality of subfragments of the same tagged long fragment comprise tag sequences from the same single bead,wherein step (b) is carried out under conditions that promote the interaction of only one tag sequence per long fragment,wherein steps (a)-(c) occur in a single vessel without physical compartmentalization of the long fragments or the beads.

View all claims
  • 2 Assignments
Timeline View
Assignment View
    ×
    ×