Multiplex amplification of short tandem repeat loci
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1. A method of simultaneously determining the alleles present in at least four short tandem repeat loci from one or more DNA samples, comprising:
- (a) obtaining at least one DNA sample to be analyzed,(b) selecting a set of at least four short tandem repeat loci of the DNA sample to be analyzed which can be amplified together, wherein the at least four loci in the set are selected from the group of loci consisting of;
D3S1539, D4S2368, D5S818, D7S820, D9S930, D10S1239, D13S317, D14S118, D14S548, D14S562, D16S490, D16S539, D16S753, D17S1298, D17S1299, D19S253, D20S481, D22S683, HUMCSF1PO, HUMTPOX, HUMTH01, HUMF13A01, HUMBFXIII, HUMLIPOL, HUMvWFA31;
(c) co-amplifying the loci in the set in a multiplex amplification reaction, wherein the product of the reaction is a mixture of amplified alleles from each of the co-amplified loci in the set; and
(d) evaluating the amplified alleles in the mixture to determine the alleles present at each of the loci analyzed in the set within the DNA sample.
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Abstract
The present invention is directed to the simultaneous amplification of multiple distinct genetic loci using PCR or other amplification systems to determine in one reaction the alleles of each of the loci contained within the multiplex.
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Citations
43 Claims
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1. A method of simultaneously determining the alleles present in at least four short tandem repeat loci from one or more DNA samples, comprising:
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(a) obtaining at least one DNA sample to be analyzed, (b) selecting a set of at least four short tandem repeat loci of the DNA sample to be analyzed which can be amplified together, wherein the at least four loci in the set are selected from the group of loci consisting of; D3S1539, D4S2368, D5S818, D7S820, D9S930, D10S1239, D13S317, D14S118, D14S548, D14S562, D16S490, D16S539, D16S753, D17S1298, D17S1299, D19S253, D20S481, D22S683, HUMCSF1PO, HUMTPOX, HUMTH01, HUMF13A01, HUMBFXIII, HUMLIPOL, HUMvWFA31; (c) co-amplifying the loci in the set in a multiplex amplification reaction, wherein the product of the reaction is a mixture of amplified alleles from each of the co-amplified loci in the set; and (d) evaluating the amplified alleles in the mixture to determine the alleles present at each of the loci analyzed in the set within the DNA sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 41, 42)
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16. A method of simultaneously determining the alleles present in three short tandem repeat loci from one or more DNA samples, comprising:
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(a) obtaining at least one DNA sample to be analyzed, (b) selecting a set of three short tandem repeat loci of the DNA sample to be analyzed which can be amplified together, wherein the set of three loci is selected from the group of sets of loci consisting of; D3S1539, D19S253, D13S317; D10S1239, D9S930, D20S481; D10S1239, D4S2368, D20S481; D10S1239, D9S930, D4S2368; D16S539, D7S820, D13S317; and D10S1239, D9S930, D13S317. (c) co-amplifying the three loci in the set in a multiplex amplification reaction, wherein the product of the reaction is a mixture of amplified alleles from each of the co-amplified loci in the set; and (d) evaluating the amplified alleles in the mixture to determine the alleles present at each of the loci analyzed in the set within the DNA sample. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24)
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25. A kit for simultaneously analyzing short tandem repeat sequences in at least three loci, comprising a container which has oligonucleotide primers for co-amplifying a set of at least three short tandem repeat loci, wherein the set of loci are selected from the sets of loci consisting of:
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D3S1539, D19S253, D13S317; D10S1239, D9S930, D20S481; D10S1239, D4S2368, D20S481; D10S1239, D9S930, D4S2368; D16S539, D7S820, D13S317; D10S1239, D9S930, D13S317; D3S1539, D7S820, D13S317, D5S818; D17S1298, D7S820, D13S317, D5S818; D20S481, D7S820, D13S317, D5S818; D9S930, D7S820, D13S317, D5S818; D10S1239, D7S820, D13S317, D5S818; D14S118, D7S820, D13S317, D5S818; D14S562, D7S820, D13S317, D5S818; D14S548, D7S820, D13S317, D5S818; D16S490, D7S820, D13S317, D5S818; D17S1299, D7S820, D13S317, D5S818; D16S539, D7S820, D13S317, D5S818; D22S683, D7S820, D13S317, D5S818; D16S753, D7S820, D13S317, D5S818; D3S1539, D19S253, D13S317, D20S481; D3S1539, D19S253, D4S2368, D20S481; D10S1239, D9S930, D4S2368, D20S481; D16S539, D7S820, D13S317, HUMvWFA31; D16S539, D7S820, D13S317, D5S818, HUMCSF1PO, HUMTPOX; D16S539, D7S820, D13S317, D5S818, HUMF13A01, HUMFESFPS; D16S539, D7S820, D13S317, D5S818, HUMCSF1PO, HUMTPOX, HUMTH01; D16S539, D7S820, D13S317, D5S818, HUMF13A01, HUMFESFPS, HUMBFXIII; D16S539, D7S820, D13S317, D5S818, HUMCSF1PO, HUMTPOX, HUMTH01, HUMvWFA31; and D16S539, D7S820, D13S317, D5S818, HUMF13A01, HUMFESFPS, HUMBFXIII, HUMLIPOL. - View Dependent Claims (26, 27, 28, 29, 30, 31, 43)
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32. A method of simultaneously determining the alleles present in at least four short tandem repeat loci from one or more DNA samples, comprising:
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(a) obtaining at least one DNA sample to be analyzed, (b) selecting a set of at least four short tandem repeat loci of the DNA sample to be analyzed which can be amplified together, wherein three of the loci in the set are D7S820, D13S317, and D5S818; (c) co-amplifying the loci in the set in a multiplex amplification reaction, wherein the product of the reaction is a mixture of amplified alleles from each of the co-amplified loci in the set; and (d) evaluating the amplified alleles in the mixture to determine the alleles present at each of the loci analyzed in the set within the DNA sample. - View Dependent Claims (33, 34, 35, 36, 37, 38, 39, 40)
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Specification