Method for selectively isolating a nucleic acid

• US 20080090733A1
• Filed: 03/13/2007
• Published: 04/17/2008
• Est. Priority Date: 12/10/1999
• Status: Abandoned Application

• Current Assignee: Generation Biotech, LLC, The Trustees of Princeton University (Princeton University)
• Original Assignee: Generation Biotech, LLC, The Trustees of Princeton University (Princeton University)
• Inventors: Cleary, Michele, Dapprich, Johannes
• Application Number:
  US11/724,043
• Publication Number:
  US 20080090733A1
• Time in Patent Office: Days
• US Class Current: 506/3
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase

Abstract

Provided are methods for selectively identifying and isolating nucleic acids in a population of nucleic acid molecules.

Claims

1-20. -20. (canceled)

21. A method for separating a polynucleotide molecule from a population of genomic DNA molecules, the method comprising:

(a) providing a population of genomic DNA molecules comprising said polynucleotide molecule, wherein said polynucleotide molecule includes a target nucleic acid sequence and a distinguishing element;

(b) contacting said population of genomic DNA molecules with a targeting element, wherein said targeting element binds specifically to said target nucleic acid sequence of said polynucleotide molecule;

(c) selectively attaching multiple separation groups to said bound targeting element, wherein attachment of separation groups occurs only if said targeting element is bound to said target nucleic acid sequence by extending said targeting element so as to include multiple separation groups;

(d) immobilizing said attached separation groups to a substrate, thereby forming an immobilized polynucleotide-targeting element-separation group complex; and

(e) removing said immobilized polynucleotide-targeting element-separation group complex from said population of genomic DNA molecules, thereby separating said polynucleotide molecule from said population of genomic DNA molecules;

wherein the distinguishing element is a polymorphism, the targeting element is an oligonucleotide that partially overlaps the distinguishing element, the separation group is an immobilizable, non-terminating nucleotide, and the 3′

-terminus of the oligonucleotide is complementary to the polymorphism.

22. The method of claim 21, wherein said separation groups are biotinylated nucleotides.

23. The method of claim 21, wherein said substrate is a particle, bead, magnetic bead, or glass surface.

24. The method of claim 21, wherein the immobilized polynucleotide-targeting element-separation group complex is topologically attached to the substrate via multiple separation groups.

25. The method of claim 21, wherein step (c) is performed by extending the 3′

terminating nucleotide by a DNA polymerase.

26. The method of claim 21, wherein the polymorphism is a single nucleotide polymorphism.