In this paper, four different applications are presented in forensic epigenetics using bisulfite modified PCR, HRM, and pyrosequencing. The first application involves the development of a multiplex amplification capable of simultaneous analysis of 4 different body fluid/cell types-blood, saliva, sperm, and vaginal epithelia. We have also developed a method for the detection of sperm high-resolution melt analysis (HRM). Two other applications involve a determination of suspect age and a biomarker for determining smoking status.
The recent development of small, single-amplicon-based benchtop systems for pyrosequencing has opened up a host of novel procedures for applications in forensic science. Pyrosequencing is a sequencing by synthesis technique, based on chemiluminescent inorganic pyrophosphate detection. This review explains the pyrosequencing workflow and illustrates the step-by-step chemistry, followed by a description of the assay design and factors to keep in mind for an exemplary assay. Existing and potential forensic applications are highlighted using this technology. Current applications include identifying species, identifying bodily fluids, and determining smoking status. We also review progress in potential applications for the future, including research on distinguishing monozygotic twins, detecting alcohol and drug abuse, and other phenotypic characteristics such as diet and body mass index. Overall, the versatility of the pyrosequencing technologies renders it a useful tool in forensic genomics.
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