Evaluation of Degradation in DNA from Males with a Quantitative Gender Typing, Endpoint PCR Multiplex

Byron C. Smith, Emily Vandegrift, Valerie Mattimore Fuller, Robert Allen

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Pages (from-to)399-408
Number of pages10
JournalJournal of Forensic Sciences
Volume60
Issue number2
DOIs
StatePublished - 1 Jan 2015

Fingerprint

Multiplex Polymerase Chain Reaction
DNA
Molecular Weight
Y Chromosome
Mitochondrial DNA
Heating
Fluorescence
Alleles

Keywords

  • Amelogenin
  • DNA degradation
  • Endpoint PCR
  • Forensic science
  • Q-TAT assay
  • Quantitative fluorescence
  • SRY

Cite this

Smith, Byron C. ; Vandegrift, Emily ; Fuller, Valerie Mattimore ; Allen, Robert. / Evaluation of Degradation in DNA from Males with a Quantitative Gender Typing, Endpoint PCR Multiplex. In: Journal of Forensic Sciences. 2015 ; Vol. 60, No. 2. pp. 399-408.
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Evaluation of Degradation in DNA from Males with a Quantitative Gender Typing, Endpoint PCR Multiplex. / Smith, Byron C.; Vandegrift, Emily; Fuller, Valerie Mattimore; Allen, Robert.

In: Journal of Forensic Sciences, Vol. 60, No. 2, 01.01.2015, p. 399-408.

Research output: Contribution to journalArticle

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