Hybridization Capture of Ancient DNA Using RNA Baits

Methods Mol Biol. 2019:1963:121-128. doi: 10.1007/978-1-4939-9176-1_13.

Abstract

The majority of DNA recovered from ancient remains is derived from organisms that colonize the remains post-mortem, such as soil microbes, or from contaminants, such as DNA from living humans. Additionally, some ancient DNA research projects aim to target specific genomic regions, such as mitochondrial genomes or variable single nucleotide polymorphisms (SNPs). To overcome the challenge of targeting specific fragments of DNA from within a complex DNA extract, methods have been developed to enrich ancient DNA extracts for target DNA relative to nontarget DNA. This chapter describes a method for target DNA enrichment that uses hybridization to biotinylated RNA baits to capture and amplify specific ancient DNA fragments from within the pool of extracted fragments.

Keywords: Ancient DNA; DNA capture; Hybridization; RNA bait.

MeSH terms

  • DNA Contamination*
  • DNA, Ancient / chemistry
  • DNA, Ancient / isolation & purification*
  • Gene Library
  • Genomics
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Nucleic Acid Hybridization / methods*
  • RNA / chemistry
  • RNA / genetics*
  • Sequence Analysis, DNA / methods

Substances

  • DNA, Ancient
  • RNA