Comparison and optimization of protocols and whole-genome capture conditions for ancient DNA samples

Biotechniques. 2024 May;76(5):216-223. doi: 10.2144/btn-2023-0107. Epub 2024 Mar 26.

Abstract

Ancient DNA (aDNA) obtained from human remains is typically fragmented and present in relatively low amounts. Here we investigate a set of optimal methods for producing aDNA data by comparing silica-based DNA extraction and aDNA library preparation protocols. We also test the efficiency of whole-genome enrichment (WGC) on ancient human samples by modifying a number of parameter combinations. We find that the Dabney extraction protocol performs significantly better than alternatives. We further observed a positive trend with the BEST library protocol indicating lower clonality. Notably, our results suggest that WGC is effective at retrieving endogenous DNA, particularly from poorly-preserved human samples, by increasing human endogenous proportions by 5x. Thus, aDNA studies will be most likely to benefit from our results.

Keywords: DNA extraction; aDNA library; ancient DNA; optimization; whole genome capture.

Publication types

  • Research Support, Non-U.S. Gov't
  • Comparative Study

MeSH terms

  • DNA, Ancient* / analysis
  • DNA, Ancient* / isolation & purification
  • Gene Library
  • Genome, Human* / genetics
  • Humans
  • Sequence Analysis, DNA / methods
  • Silicon Dioxide / chemistry

Substances

  • DNA, Ancient
  • Silicon Dioxide