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Hair root sonication washing impact on nuclei counts.

Stephanie M Betts1,2, Ivette A Espinoza Quiroga1,2, Linda M Otterstatter3

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Sonication washing of hair roots may reduce nuclei for DNA extraction. For nuclear DNA analysis only, consider less aggressive methods like vortexing and ethanol rinsing to preserve nuclei.

Keywords:
DAPI stainingfluorescence microscopyforensic hair analysishair root nucleinuclear DNA typing

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Area of Science:

  • Forensic Science
  • Molecular Biology
  • Genetics

Background:

  • Sonication washing is used in DNA extraction protocols for hair samples.
  • This step was previously considered necessary for both mitochondrial (mt) and nuclear (nu) DNA analyses.

Purpose of the Study:

  • To assess the impact of sonication washing on nuclei persistence in hair roots.
  • To determine if sonication affects the number of nuclei available for DNA extraction.

Main Methods:

  • Scalp hair roots were stained with 4', 6-diamidino-2-phenylindole (DAPI).
  • Visible nuclei were counted in both unwashed and sonication-washed hair roots.
  • Hair roots with varying initial nuclei counts (<50 and >100) were analyzed.

Main Results:

  • Sonication washing may reduce the number of nuclei in hair roots, particularly those with fewer initial nuclei.
  • Challenges in nuclei counting included hair root orientation, morphology, and tissue folding.
  • A reduction in nuclei could impact DNA extraction yield and short tandem repeat (STR) typing.

Conclusions:

  • Sonication washing might deplete nuclei essential for DNA extraction and STR typing.
  • Laboratories performing only nuclear DNA analysis can opt for gentler washing methods, like low-speed vortexing and ethanol rinsing.