Ancient DNA pp 75-83 | Cite as

A Method for Single-Stranded Ancient DNA Library Preparation

  • Marie-Theres GansaugeEmail author
  • Matthias Meyer
Part of the Methods in Molecular Biology book series (MIMB, volume 1963)


Genomic library preparation from highly degraded DNA is more efficient when library molecules are prepared separately from the complementary strands of DNA fragments. We describe a protocol in which libraries are constructed from single DNA strands in a three-step procedure: single-stranded ligation of the first adapter with T4 DNA ligase in the presence of a splinter oligonucleotide, copying of the DNA strand with a proofreading polymerase, and blunt-end ligation of the second double-stranded adapter with T4 DNA ligase.

Key words

NGS Library preparation Ancient DNA Single-stranded ligation T4 DNA ligase 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Evolutionary GeneticsMax Planck Institute for Evolutionary AnthropologyLeipzigGermany

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