Pretreatment: Removing DNA Contamination from Ancient Bones and Teeth Using Sodium Hypochlorite and Phosphate

Methods Mol Biol. 2019:1963:15-19. doi: 10.1007/978-1-4939-9176-1_2.

Abstract

DNA isolated from ancient bones and teeth comprises a mixture of microbial contamination and DNA from the organism under study. In addition, analyses of ancient human remains are often complicated by contamination with present-day human DNA, which can be introduced during excavation and subsequent handling of the specimens. In most cases, the relative abundance of contaminant DNA is much greater than that of the target organism. Here we present two techniques for reducing the proportion of contaminant DNA in bones and teeth. The first and most efficient technique uses a sodium hypochlorite (bleach) pretreatment to destroy contaminant DNA that may be bound or otherwise attached to the surface of bone/tooth powder. The second, less destructive pretreatment uses a phosphate buffer to release surface-bound DNA.

Keywords: Ancient DNA; DNA contamination; DNA extraction; Pretreatment; Sodium hypochlorite (bleach); Sodium phosphate.

MeSH terms

  • Animals
  • Bone and Bones / metabolism*
  • DNA / analysis*
  • DNA / chemistry
  • DNA / isolation & purification*
  • DNA Contamination*
  • Decontamination
  • Neanderthals
  • Phosphates / chemistry*
  • Polymerase Chain Reaction
  • Sodium Hypochlorite / chemistry*
  • Specimen Handling / methods
  • Tooth / metabolism*

Substances

  • Phosphates
  • DNA
  • Sodium Hypochlorite