Patent ID: 8153372

Claim:
A method for determining, in a sample, using a single multiplex PCR reaction, each of: a quantity of genomic DNA from a male donor and a quantity of genomic DNA from a female donor, a ratio between said quantity of genomic DNA from said male donor and said quantity of genomic DNA from said female donor, extent of genomic DNA degradation, and presence of PCR inhibitors, comprising the steps of i) creating a reaction mix by combining an aliquot of said sample and DNA encoding a non-human reporter gene; ii) amplifying said reaction mix in a single multiplex PCR reaction using a plurality of primer sets comprising: a) a first primer set comprising synthetic oligonucleotide primers directed against human male and female amelogenin genetic loci; b) a second primer set comprising synthetic oligonucleotide primers directed against a human Y-chromosome specific gene or a human X-chromosome specific gene, or both; wherein amplicons produced from said human Y-chromosome specific gene and said human X-chromosome specific gene are shorter than amplicons produced from said human male and female amelogenin genetic loci; and c) a third primer set comprising synthetic oligonucleotide primers directed against said non-human reporter gene; iii) detecting PCR amplicons produced in said step of amplifying; and iv) making an assessment of: a) said quantity of genomic DNA from a male donor and said quantity of genomic DNA from a female donor, b) said ratio between said quantity of genomic DNA from said male donor and said quantity of genomic DNA from said female donor, c) said extent of genomic DNA degradation, and d) said presence of PCR inhibitors in said sample based on quantities of non-human PCR amplicons detected in said detecting step, wherein said step of making an assessment of said extent of genomic degradation includes a step of i. calculating a ratio of: PCR amplicons produced by amplifying said human Y-chromosome specific gene and said human male amelogenin genetic locus; and PCR amplicons produced by amplifying said human X-chromosome specific gene and said human female amelogenin genetic locus.