Patent ID: 9040256
Filing Date: 2015-05-26
Classification: C12N,C12P,C12Q

Abstract:
1. A method for parallel sequencing of a target DNA in a sample, the method comprising: (a) providing: (b) incubating the target DNA with the plurality of transposome complexes under conditions whereby the target DNA is fragmented into double-stranded DNA fragments comprising a transferred strand from the transposon end compositions joined to the 5′ ends of the double-stranded DNA fragments, thereby generating 5′-tagged DNA fragments; (c) extending the 3′ ends of the 5′-tagged DNA fragments; (d) non-selectively amplifying the extended 5′-tagged DNA fragments under conditions that do not intentionally select for or against amplification of particular 5′-tagged DNA fragments based on their target DNA sequences to generate a representative library of amplified DNA fragments having a population that is representative of the target DNA from which the fragments were generated, wherein each amplified DNA fragment comprises: (e) capturing a plurality of the amplified DNA fragments on a surface through the capture tag; (f) providing a plurality of sequencing primers that anneal to a portion of each captured DNA fragment; and (g) simultaneously sequencing the captured fragments in a single multiplex format to determine the nucleotide sequences of the DNA fragments captured on the surface in parallel.