Maximizing efficiency in sedimentary ancient DNA analysis: a novel extract pooling approach

Victoria Oberreiter, Pere Gelabert (Korresp. Autor*in), Florian Brück, Stefan Franz, Evelyn Zelger, Sophie Szedlacsek, Olivia Cheronet, Fernanda Tenorio Cano, Florian Exler, Brina Zagorc, Ivor Karavanić, Marko Banda, Boris Gasparyan, Lawrence Guy Straus, Manuel R Gonzalez Morales, John Kappelman, Mareike Stahlschmidt, Thomas Rattei, Stephan M Kraemer, Susanna Sawyer (Korresp. Autor*in)Ron Pinhasi (Korresp. Autor*in)

Veröffentlichungen: Beitrag in FachzeitschriftArtikelPeer Reviewed

Abstract

In the last few decades, the field of ancient DNA has taken a new direction towards using sedimentary ancient DNA (sedaDNA) for studying human and mammalian population dynamics as well as past ecosystems. However, the screening of numerous sediment samples from archaeological sites remains a time-consuming and costly endeavor, particularly when targeting hominin DNA. Here, we present a novel high-throughput method that facilitates the fast and efficient analysis of sediment samples by applying a pooled testing approach. This method combines multiple extracts, enabling early parallelization of laboratory procedures and effective aDNA screening. Pooled samples with detectable aDNA signals undergo detailed analysis, while empty pools are discarded. We have successfully applied our method to multiple sediment samples from Middle and Upper Paleolithic sites in Europe, Asia, and Africa. Notably, our results reveal that an aDNA signal remains discernible even when pooled with four negative samples. We also demonstrate that the DNA yield of double-stranded libraries increases significantly when reducing the extract input, potentially mitigating the effects of inhibition. By embracing this innovative approach, researchers can analyze large numbers of sediment samples for aDNA preservation, achieving significant cost reductions of up to 70% and reducing hands-on laboratory time to one-fifth.

OriginalspracheEnglisch
Aufsatznummer19388
Seiten (von - bis)19388
FachzeitschriftScientific Reports
Jahrgang14
Ausgabenummer1
DOIs
PublikationsstatusVeröffentlicht - 20 Aug. 2024

ÖFOS 2012

  • 106018 Humanbiologie

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