Patent ID: 9725765
Date: 2017-08-08
CPC Classifications: C12Q,Y02A

Claim:
1. A method for analyzing a plurality of target polynucleotides, the method comprising: (a) providing a sample comprising a plurality of target polynucleotide fragments, wherein each target polynucleotide fragment has a length of 5 kb to 40 kb; (b) performing a size selection on the plurality of target polynucleotide fragments to select 5-7 kb fragments; (c) ligating amplification adaptors at each end of at least a portion of the 5-7 kb fragments to produce a library of amplifiable fragments, wherein the adaptors serve as primer binding sites; (d) quantitating via qPCR to establish a number of amplifiable fragments in the library of amplifiable fragments; (e) determining a dilution factor based on the number of amplifiable fragments in the library of amplifiable fragments; (f) diluting and partitioning the library into a plurality of partitions based on the dilution factor such that each partition comprises, on average, a unique subset of 500 or more of the amplifiable fragments; (g) amplifying via polymerase chain reaction the partitioned amplifiable fragments; (h) subjecting the partitions to polynucleotide fragmentation thereby generating a set of smaller fragments; and (i) obtaining partial or complete sequence information from at least a subset of the smaller fragments.