Patent ID: 9481909
Filing Date: 2016-11-01
CPC Classification: C12Q

Claim Text:
1. A method comprising: a. conducting a multiplex real-time Polymerase Chain Reaction (PCR) on a sample, comprising amplifying a human nuclear DNA target that is not a polymorphic short tandem repeat (STR), a human Y chromosome target and an internal PCR control (IPC), wherein the sequence of each primer of a primer pair for the human nuclear DNA target differs from the sequence of each primer of a primer pair for the human Y chromosome target and the IPC is not homologous to mammalian DNA; b. detecting the presence or absence of the amplified human nuclear DNA target that is not a polymorphic STR, amplified human Y chromosome target and amplified IPC by fluorescence; c. determining the accurate quantity of the amplified human nuclear DNA target that is not a polymorphic STR, the human Y chromosome target, and the IPC, wherein the amount of amplified human nuclear DNA target that is not a polymorphic STR and the amount of amplified human Y chromosome target correspond to a quantitative value of the amount of the amplified targets respectively and the amount of the amplified IPC indicates the accuracy of the quantitation, and thereby determining the quantity of amplifiable DNA in the sample, wherein the length of the amplified human nuclear DNA target that is not a polymorphic STR and the human Y chromosome target differs by no more than 20 nucleotides from each other; d. determining the ratio of human nuclear DNA target that is not a polymorphic STR to the human Y chromosome target DNA, wherein the amount of the human Y chromosome target DNA corresponds to the amount of human male DNA in the sample; e. determining the level of PCR inhibitors in the sample comprising analysis of the amplification of the IPC, the human nuclear DNA target that is not a polymorphic STR and the human male DNA target in the presence of known amounts of PCR inhibitors; f. adjusting the amount of the sample required to perform a STR genotyping assay based on the quantity of amplifiable DNA; g. selecting a STR genotyping assay system to be used for STR analysis based on the amount of human male DNA present in the sample and the level of PCR inhibitors present in the sample; and h. performing a multiplex STR assay on the adjusted amount of the sample using the STR genotyping assay system selected.