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Evaluation of Degradation in DNA from Males with a Quantitative Gender Typing, Endpoint PCR Multiplex
Author(s) -
Smith Byron C.,
Vandegrift Emily,
Fuller Valerie Mattimore,
Allen Robert W.
Publication year - 2015
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/1556-4029.12682
Subject(s) - amplicon , mitochondrial dna , dna , microbiology and biotechnology , multiplex polymerase chain reaction , nuclear dna , multiple displacement amplification , multiplex , biology , polymerase chain reaction , degradation (telecommunications) , genomic dna , chemistry , genetics , gene , dna extraction , telecommunications , computer science
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 DNA s 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.