Double-Stranded Library Preparation for Ancient and Other Degraded Samples.

High-throughput sequencing (HTS) allows fast and cost-efficient sequencing of ancient DNA (aDNA) without prior information about what sequences should be targeted. One necessary step for HTS is the preparation of a sequencing library. Commercial kits are available for this purpose, but many of these are not suitable for aDNA or other types of damaged DNA. Here, we outline a protocol for HTS library preparation that is optimized for ancient DNA. We report the library conversion rate for a range of input template and adapter concentrations. Our results show that the protocol performs at a high efficiency.

[1]  Jesse Dabney,et al.  Length and GC-biases during sequencing library amplification: a comparison of various polymerase-buffer systems with ancient and modern DNA sequencing libraries. , 2012, BioTechniques.

[2]  M. Meyer,et al.  Single-stranded DNA library preparation for the sequencing of ancient or damaged DNA , 2013, Nature Protocols.

[3]  I. Lehman DNA ligase: structure, mechanism, and function. , 1974, Science.

[4]  Swapan Mallick,et al.  Complete Genomes Reveal Signatures of Demographic and Genetic Declines in the Woolly Mammoth , 2015, Current Biology.

[5]  V. Radchuk,et al.  Two species of Southeast Asian cats in the genus Catopuma with diverging histories: an island endemic forest specialist and a widespread habitat generalist , 2016, Royal Society Open Science.

[6]  Matthias Meyer,et al.  Illumina sequencing library preparation for highly multiplexed target capture and sequencing. , 2010, Cold Spring Harbor protocols.

[7]  M. Hofreiter,et al.  Massive influence of DNA isolation and library preparation approaches on palaeogenomic sequencing data , 2016, bioRxiv.

[8]  Adrian W. Briggs,et al.  Analysis of one million base pairs of Neanderthal DNA , 2006, Nature.

[9]  R. Vasconcelos,et al.  Phylogeography of the Small Indian Civet and Origin of Introductions to Western Indian Ocean Islands , 2016, The Journal of heredity.

[10]  Arne Ludwig,et al.  The future of ancient DNA: Technical advances and conceptual shifts , 2015, BioEssays : news and reviews in molecular, cellular and developmental biology.

[11]  Christina Backes,et al.  An integer linear programming approach for finding deregulated subgraphs in regulatory networks , 2011, Nucleic acids research.

[12]  S. Turvey,et al.  Evolutionary History of the Nesophontidae, the Last Unplaced Recent Mammal Family. , 2016, Molecular biology and evolution.

[13]  Edward M. Rubin,et al.  Genomic Sequencing of Pleistocene Cave Bears , 2005, Science.

[14]  Adrian W. Briggs,et al.  Preparation of next-generation sequencing libraries from damaged DNA. , 2012, Methods in molecular biology.

[15]  H. Gan,et al.  Phylogeography of red muntjacs reveals three distinct mitochondrial lineages , 2017, BMC Evolutionary Biology.

[16]  Johanna L A Paijmans,et al.  Analysis of Whole Mitogenomes from Ancient Samples. , 2015, Methods in molecular biology.

[17]  Martin Kircher,et al.  Double indexing overcomes inaccuracies in multiplex sequencing on the Illumina platform , 2011, Nucleic acids research.