Double-Stranded Library Preparation for Ancient and Other Degraded Samples

Kirstin Henneberger; Axel Barlow; Johanna L A Paijmans

doi:10.1007/978-1-4939-9176-1_8

Double-Stranded Library Preparation for Ancient and Other Degraded Samples

Methods Mol Biol. 2019:1963:65-73. doi: 10.1007/978-1-4939-9176-1_8.

Authors

Kirstin Henneberger¹, Axel Barlow¹, Johanna L A Paijmans²

Affiliations

¹ Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany.
² Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany. paijmans.jla@gmail.com.

PMID: 30875045
DOI: 10.1007/978-1-4939-9176-1_8

Abstract

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.

Keywords: Ancient DNA; Degraded DNA; Double-stranded library preparation; Historical DNA; Illumina sequencing.

MeSH terms

DNA / genetics*
DNA Damage*
DNA, Ancient / analysis*
DNA, Ancient / chemistry
DNA, Ancient / isolation & purification
Gene Library*
High-Throughput Nucleotide Sequencing / methods*
Sequence Analysis, DNA / methods*

Substances

DNA, Ancient
DNA