METHODS FOR RAPID FORENSIC DNA ANALYSIS
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Abstract
The present invention provides methods and primer pairs for rapid, high-resolution forensic analysis of DNA and STR-typing by using amplification and mass spectrometry, determining the molecular masses and calculating base compositions of amplification products and comparing the molecular masses with the molecular masses of theoretical amplicons indexed in a database.
158 Citations
136 Claims
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1-115. -115. (canceled)
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116. A method for STR typing comprising:
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a) amplifying a nucleic acid from a sample with an oligonucleotide primer pair comprising a forward and a reverse primer, each between 13 and 40 nucleobases in length, wherein said forward primer and is configured to hybridize within a first conserved region of said nucleic acid and said reverse primer is configured to hybridize within a second conserved region of said nucleic acid, wherein said first and said second conserved regions flank a variable nucleic acid region comprising a STR locus, wherein said amplifying generates at least one amplification product that is between about 45 and about 200 nucleotides in length; b) determining the molecular mass of at least one strand of said at least one amplification product by mass spectrometry; and c) comparing said molecular mass to a molecular mass database comprising a plurality of molecular masses of a plurality of STR-identifying amplification products indexed to said oligonucleotide primer pair and to a reference allele corresponding to said STR locus, wherein a match between said determined molecular mass and a molecular mass comprised in said molecular mass database identifies an STR allele in said sample. - View Dependent Claims (117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130)
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131. A purified oligonucleotide primer pair configured to generate an amplification product that is between about 45 and about 200 nucleotides in length comprising a forward and a reverse primer, each between 13 and 40 nucleobases in length
wherein said forward primer is configured to hybridize within a first conserved region of a nucleic acid and said reverse primer is configured to hybridize within a second conserved region of said nucleic acid, wherein said first and second conserved regions flank a variable nucleic acid region comprising an STR locus selected from the group consisting of: - VWA, TPOX, THO1, FGA, D21S11, D18S51, D16S539, D13S317, D8S1179, D7S820, D5S818, D3S, and CSF1PO,
said primer pair comprising at least 70%, at least 80%, at least 90%, at least 95% or at least 100% sequence identity with an oligonucleotide primer pair selected from the group consisting of SEQ ID NOs;
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104. - View Dependent Claims (132, 133)
- VWA, TPOX, THO1, FGA, D21S11, D18S51, D16S539, D13S317, D8S1179, D7S820, D5S818, D3S, and CSF1PO,
- 134. A kit for forensic analysis comprising at least two oligonucleotide primer pairs configured to generate one or more amplification products between about 45 and about 200 nucleotides in length comprising a forward and a reverse primer, wherein the first of said at least two oligonucleotide primer pairs comprises a first forward primer configured to hybridize within a first conserved region of said nucleic acid and a first reverse primer configured to hybridize within a second conserved region of said nucleic acid, wherein said first and second conserved regions flank a variable nucleic acid region comprising vWR, and wherein the second of said at least two oligonucleotide primer pairs is configured to hybridize within an AMEL locus or to regions flanking an AMEL locus.
Specification