Year: 2025
Type: article
Abstract: Large-scale DNA screening of palaeontological and archaeological collections remains a limiting and costly factor for ancient DNA studies. Several DNA extraction protocols are routinely used in ancient DNA laboratories and have even been automated on robotic platforms. Robots offer a solution for high-throughput screening but the costs, as well as necessity for trained technicians and engineers, can be prohibitive for some laboratories. Here, we present a high-throughput alternative to robot-based ancient DNA extraction using a 96-column plate. When compared to routine single MinElute columns, we retrieved highly similar endogenous DNA contents, an important metric in ancient DNA screening. Mitogenomes with a coverage depth greater than 0.1× could be generated and allowed for taxonomic assignment. However, average fragment lengths, DNA damage and library complexities significantly differed between methods but these differences became nonsignificant after modification of our library purification protocol. Our high-throughput extraction method allows generation of 96 extracts within approximately 4 hours of laboratory work while bringing the cost down by ~39% compared to using single columns. Additionally, we formally demonstrate that the addition of Tween-20 during the elution step results in higher complexity libraries, thereby enabling higher genome coverage for the same sequencing effort.
Source: Molecular Ecology Resources
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $5,250
Awards: G1322033128, G230487938, G247595552, DeepTime Palaeogenomics - Tracing species evolution using ancient genomes sampled on a million-year time scale, G4066480865, G4353964202, G5894106689, G6832510969, G7054268934, G7230600129, G808781930, MAMMOTH ARCTIC PATHS (MAP): Interdisciplinary Research into the Movement Ecology and Biogeography of an Iconic Arctic Animal through Environmental Change., G8981540644
OpenAlex ID: W4407225565