Use of bleach to eliminate contaminating DNA from the surface of bones and teeth

The extraction of DNA from archaeological or forensic skeletal remains can provide quite powerful data for analysis, but is plagued by a unique set of methodological problems. One of the most important methodological problems to overcome in such analyses is the presence of modern contamination on th...

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Bibliographic Details
Published in:Forensic science international 2005-11, Vol.154 (1), p.53-61
Main Authors: Kemp, Brian M., Smith, David Glenn
Format: Article
Language:English
Subjects:
Analysis
Ancient DNA
Biological and medical sciences
Bleaching materials
Contamination
Contamination (Technology)
Deoxyribonucleic acid
Disinfectants - administration & dosage
DNA
DNA - chemistry
DNA - isolation & purification
DNA testing
Forensic Anthropology - methods
Forensic medicine
Forensic sciences
Fossils
General aspects
Genetic aspects
Human skeleton
Humans
Hydroxyapatite
Immersion
Investigative techniques, diagnostic techniques (general aspects)
Medical sciences
Methods
Polymerase Chain Reaction
Public health. Hygiene
Public health. Hygiene-occupational medicine
R&D
Research & development
Ribs - chemistry
Skeletal remains
Sodium hypochlorite
Sodium Hypochlorite - administration & dosage
Studies
Teeth
Time Factors
Tooth - chemistry
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Summary:The extraction of DNA from archaeological or forensic skeletal remains can provide quite powerful data for analysis, but is plagued by a unique set of methodological problems. One of the most important methodological problems to overcome in such analyses is the presence of modern contamination on the surfaces of bones and teeth, which can lead to false positives and erroneous results unless it is removed before DNA extraction is initiated. Ancient DNA (aDNA) researchers and forensic scientists have employed a number of techniques to minimize such contamination. One such technique is the use of bleach (sodium hypochlorite—NaOCl) to “destroy” contaminating DNA. However, a consensus on the optimum concentration of sodium hypochlorite to be used and the amount of time the bone or tooth should be exposed to it has not emerged. The present study systematically approaches the issue by introducing contamination to ancient bones (from ∼500 BP) and determining which of several sodium hypochlorite treatments best eliminates surface contamination. The elimination of surface contamination from bone requires immersion in at least 3.0% (w/v) sodium hypochlorite (approximately equal parts of commercial bleach and water) for at least 15 min. Endogenous DNA proved to be quite stable to even extreme sodium hypochlorite treatments (6% for 21 h), suggesting that DNA adsorbs to hydroxyapatite in the bone and that this process facilitates the preservation of DNA in ancient skeletal remains.
ISSN:0379-0738
1872-6283
DOI:10.1016/j.forsciint.2004.11.017