Methods For Rapid Forensic Analysis Of Mitochondrial DNA

• US 20090125245A1
• Filed: 03/17/2008
• Published: 05/14/2009
• Est. Priority Date: 05/25/2004
• Status: Active Grant

• Current Assignee: Ibis Biosciences Incorporated, Florida Turbine Technologies, Inc. (Kratos Defense & Security Solutions Incorporated)
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: White, Neill, Hofstadler, Steven A., Hall, Thomas A., Blyn, Lawrence B., Eshoo, Mark W., Ecker, David J., Samant, Vivek
• Granted Patent: US 8,407,010 B2
• Time in Patent Office: Days
• US Class Current: 702/20
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/683 involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
• CPC Class Codes: C12Q 1/6846 Common amplification features
• CPC Class Codes: C12Q 2521/301 Endonuclease
• CPC Class Codes: C12Q 2565/627 being a mass spectrometer
• CPC Class Codes: G01N 27/447 using electrophoresis
• CPC Class Codes: G01N 27/623 combined with mass spectrometry
• CPC Class Codes: G16B 50/00 ICT programming tools or database systems specially adapted for bioinformatics
• CPC Class Codes: G16B 50/30 Data warehousing; Computing architectures

Abstract

The present invention provides methods for rapid forensic analysis of mitochondrial DNA by amplification of a segment of mitochondrial DNA containing restriction sites, digesting the mitochondrial DNA segments with restriction enzymes, determining the molecular masses of the restriction fragments and comparing the molecular masses with the molecular masses of theoretical restriction digests of known mitochondrial DNA sequences stored in a database.

Claims

48-1. The method of claim 1 wherein said amplifying step comprises polymerase chain reaction.

64. A primer pair wherein each member of the pair has at least 70% sequence identity with the sequence of the corresponding member of any one of the following primer pair sequences:

SEQ ID NOs;

8;

9, 10;

11, 12;

13, 12;

14, 12;

15, 16;

17, 18;

19, 20;

21, 22;

23, 24;

25, 26;

27, 28;

29, 30;

31, 32;

33, 34;

35, 36;

37, 38;

39, 40;

41, 42;

43, 44;

45, 42;

46, 47;

48, 18;

49, 50;

51, 22;

52, 53;

54, 55;

56, 57;

29, 58;

31, 59;

60, 61;

62, 63;

39, 40;

64, 65;

66, 67;

68, 69;

70, 12;

68, 12;

70, 67;

15, 71;

70, 69;

15, and 69;

68.

65. The primer pair of claim 64 comprising at least one modified nucleobase.

66. The primer pair of claim 65 wherein the modified nucleobase is 5-propynylcytidine or 5-propynyluridine.

67. An isolated mitochondrial DNA amplicon comprising a segment of mitochondrial DNA produced by the process of amplification of a sample of mitochondrial DNA with the pair of primers of claim 64.