PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly

PeerJ

article

Maxime Borry¹  Alexander Hübner¹  Adam B. Rohrlach³  Christina Warinner¹ 

[1] Microbiome Sciences Group, Max Planck Institute for the Science of Human History, Department of Archaeogenetics;Faculty of Biological Sciences, Friedrich-Schiller Universität Jena;Population Genetics Group, Max Planck Institute for the Science of Human History, Department of Archaeogenetics;ARC Centre of Excellence for Mathematical and Statistical Frontiers, The University of Adelaide;Department of Anthropology, Harvard University

关键词: metagenomics;    aDNA;    ancient DNA;    assembly;    damage;    de novo;    automated;   

DOI  :  10.7717/peerj.11845

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

DNA de novo assembly can be used to reconstruct longer stretches of DNA (contigs), including genes and even genomes, from short DNA sequencing reads. Applying this technique to metagenomic data derived from archaeological remains, such as paleofeces and dental calculus, we can investigate past microbiome functional diversity that may be absent or underrepresented in the modern microbiome gene catalogue. However, compared to modern samples, ancient samples are often burdened with environmental contamination, resulting in metagenomic datasets that represent mixtures of ancient and modern DNA. The ability to rapidly and reliably establish the authenticity and integrity of ancient samples is essential for ancient DNA studies, and the ability to distinguish between ancient and modern sequences is particularly important for ancient microbiome studies. Characteristic patterns of ancient DNA damage, namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate ancient samples and sequences, but existing tools for inspecting and filtering aDNA damage either compute it at the read level, which leads to high data loss and lower quality when used in combination with de novo assembly, or require manual inspection, which is impractical for ancient assemblies that typically contain tens to hundreds of thousands of contigs. To address these challenges, we designed PyDamage, a robust, automated approach for aDNA damage estimation and authentication of de novo assembled aDNA. PyDamage uses a likelihood ratio based approach to discriminate between truly ancient contigs and contigs originating from modern contamination. We test PyDamage on both on simulated aDNA data and archaeological paleofeces, and we demonstrate its ability to reliably and automatically identify contigs bearing DNA damage characteristic of aDNA. Coupled with aDNA de novo assembly, Pydamage opens up new doors to explore functional diversity in ancient metagenomic datasets.

【 授权许可】

CC BY