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Complexity management of genomic DNA

  • US 8,492,121 B2
  • Filed: 06/29/2011
  • Issued: 07/23/2013
  • Est. Priority Date: 10/27/1999
  • Status: Expired due to Term
First Claim
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1. A method for selective amplification of a nucleic acid sample, the method comprising:

  • fragmenting the nucleic acid sample using a first restriction enzyme and a second restriction enzyme and producing a digested nucleic acid sample comprising fragments;

    ligating a double stranded first adaptor and a double stranded second adaptor to the fragments and producing ligated fragments, wherein the 3′

    end of a first strand of the first adaptor is blocked from ligating to the 5′

    ends of the fragments such that only a second strand of the first adaptor can ligate to the fragments, and wherein the 5′

    end of a first strand of the second adaptor is blocked from ligating to the 3′

    ends of the fragments such that only a second strand of the second adaptor can ligate to the fragments, wherein a subset of the ligated fragments comprises ligated fragments having a single strand which is ligated to the first adaptor at its 3′

    end and the second adaptor at its 5′

    end; and

    subjecting the ligated fragments to amplification conditions, and selectively and exponentially amplifying the ligated fragments having a single strand which is ligated to the first adaptor at its 3′

    end and the second adaptor at its 5′

    end, thereby selectively amplifying the nucleic acid sample.

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