Methods for rapid forensic analysis of mitochondrial DNA and characterization of mitochondrial DNA heteroplasmy
First Claim
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1. A method of identifying a nucleic acid sequence from a human sample, comprising:
- a) contacting nucleic acid from said human sample with two or more oligonucleotide primers that hybridize to sequence regions of said nucleic acid that are conserved among different bioagents, wherein said conserved sequence regions flank a variable sequence region to produce an amplification product;
b) determining the base composition of said amplification product wherein said base composition of said amplification product comprises identification of the number of A residues, C residues, T residues, G residues, U residues, analogs thereof and/or mass tag residues thereof and wherein said base composition is determined without sequencing said amplification product; and
c) identifying said nucleic acid sequence from said human sample by comparing said determined base composition to a database of base compositions from a plurality of different bioagents wherein said base composition of said amplification product comprises identification of the number but not the nucleic acid sequence order of A residues, C residues, T residues, G residues, U residues, analogs thereof and/or mass tag residues thereof.
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Abstract
The present invention provides methods for rapid forensic analysis of mitochondrial DNA and methods for characterizing heteroplasmy of mitochondrial DNA, which can be used to assess the progression of mitochondrial diseases.
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Citations
71 Claims
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1. A method of identifying a nucleic acid sequence from a human sample, comprising:
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a) contacting nucleic acid from said human sample with two or more oligonucleotide primers that hybridize to sequence regions of said nucleic acid that are conserved among different bioagents, wherein said conserved sequence regions flank a variable sequence region to produce an amplification product; b) determining the base composition of said amplification product wherein said base composition of said amplification product comprises identification of the number of A residues, C residues, T residues, G residues, U residues, analogs thereof and/or mass tag residues thereof and wherein said base composition is determined without sequencing said amplification product; and c) identifying said nucleic acid sequence from said human sample by comparing said determined base composition to a database of base compositions from a plurality of different bioagents wherein said base composition of said amplification product comprises identification of the number but not the nucleic acid sequence order of A residues, C residues, T residues, G residues, U residues, analogs thereof and/or mass tag residues thereof. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 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, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71)
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Specification