Modified DOP-PCR for improved STR typing of degraded DNA from human skeletal remains and bloodstains

Angie Ambers, Meredith Turnbough, Robert Benjamin, Harrell Gill-King, Jonathan King, Antti Sajantila, Bruce Budowle

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Forensic and ancient DNA samples often are damaged and in limited quantity as a result of exposure to harsh environments and the passage of time. Several strategies have been proposed to address the challenges posed by degraded and low copy templates, including a PCR based whole genome amplification method called degenerate oligonucleotide-primed PCR (DOP-PCR). This study assessed the efficacy of four modified versions of the original DOP-PCR primer that retain at least a portion of the 5' defined sequence and alter the number of bases on the 3' end. The use of each of the four modified primers resulted in improved STR profiles from environmentally-damaged bloodstains, contemporary human skeletal remains, American Civil War era bone samples, and skeletal remains of WWII soldiers over those obtained by previously described DOP-PCR methods and routine STR typing. Additionally, the modified DOP-PCR procedure allows for a larger volume of DNA extract to be used, reducing the need to concentrate the sample and thus mitigating the effects of concurrent concentration of inhibitors.

Original languageEnglish
Pages (from-to)7-12
Number of pages6
JournalLegal Medicine
Volume18
DOIs
StatePublished - 1 Jan 2016

Keywords

  • Bloodstains
  • DOP-PCR
  • Degraded DNA
  • Human skeletal remains
  • Whole genome amplification

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