摘要:
A comprehensive set of human specific, target specific, multiplex PCR assays for DNA quantitation is provided. Our duplex qPCR for nDNA/mtDNA had a linear quantitation range of 100 ng to 1 pg, and our triplex qPCR assay for nDNA/mtDNA/male Y DNA had a linear range of 100 ng to 0.1 ng. Human-specificity was demonstrated by the accurate detection of 0.05% and 5% human DNA, respectively, from a complex source of starting templates. Target-specificity was confirmed by the lack of cross-amplification among targets. A high throughput alternative for human gender determination was also developed by multiplexing the male Y primer/probe set with an X chromosome based system. Background cross-amplification with DNA templates derived from fourteen other species was negligible aside from the male Y assay which produced spurious amplifications from other non-human primate templates. Mainstream application of these assays will undoubtedly benefit forensic genomics.
摘要:
A method for determining gender from a human DNA sample. The loci of Alu element insertion is selected, amplified and evaluated in terms of size of the fragment. The gender assay utilizes AluSTXa for the X chromosome, AluSTYa for the Y chromosome, or both AluSTXa and AluSTYa, to reduce the possibility of error to a negligible quantity. The inserted chromosome yields a large fragment when the homologous region is amplified. The males are distinguished as having two DNA amplicons present, while females have only a single amplicon. The kit adapted for carrying out the method includes a pair of primers to amplify the locus and optionally polymerase chain reaction regents.
摘要:
A method for determining gender from a human DNA sample. The loci of Alu element insertion is selected, amplified and evaluated in terms of size of the fragment. The gender assay utilizes AluSTXa for the X chromosome, AluSTYa for the Y chromosome, or both AluSTXa and AluSTYa, to reduce the possibility of error to a negligible quantity. The inserted chromosome yields a large fragment when the homologous region is amplified. The males are distinguished as having two DNA amplicons present, while females have only a single amplicon. The kit adapted for carrying out the method includes a pair of primers to amplify the locus and optionally polymerase chain reaction regents.