title: Evaluation of degradation in DNA from males with a quantitative gender typing, endpoint PCR multiplex.
abstract: Evidentiary samples submitted to a forensic DNA laboratory occasionally yield DNA that is degraded. Samples of intact chromosomal DNA (both nuclear and mitochondrial) were subjected to a heating protocol to induce DNA degradation. The DNAs were then analyzed using a multiplex PCR assay that amplifies targets of low and high molecular weight on the X/Y and mitochondrial chromosomes. If degradation is random, the amplification of larger DNA targets should be more adversely affected by degradation than smaller targets. In nuclear and mitochondrial DNA from a male donor, exhibiting degradation, DNA quantity estimates based upon higher molecular weight amplicons (HMW) are significantly lower than estimates made using low molecular weight (LMW) Q-TAT amplicons. DNA degradation estimated using this approach correlated well with actual fluorescence associated with HMW and LMW STR alleles amplified from the same genomic DNA templates. Q-TAT is thus useful not only as a quantitation tool, but also as an indicator of template degradation.
authors.affiliation: Forensic Laboratory, Tulsa Police Department, 1111 W. 17th Street, Building E, 2nd Floor, Tulsa, OK.
authors.forename: Byron C
authors.initials: BC
authors.lastname: Smith
authors.forename: Emily
authors.initials: E
authors.lastname: Vandegrift
authors.forename: Valerie Mattimore
authors.initials: VM
authors.lastname: Fuller
authors.forename: Robert W
authors.initials: RW
authors.lastname: Allen
mesh_terms: D004247:DNA / Q000032:analysis*; D053836:DNA Degradation, Necrotic; D016172:DNA Fingerprinting / Q000295:instrumentation*; D017931:DNA Primers; D019075:Electrophoresis, Capillary; D006801:Humans; D008297:Male; D008970:Molecular Weight; D060885:Multiplex Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D004247:DNA
keywords: DNA degradation; Q-TAT assay; SRY; amelogenin; endpoint PCR; forensic science; quantitative fluorescence