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Ancient DNA pp 85-92 | Cite as

Sequencing Library Preparation from Degraded Samples for Non-illumina Sequencing Platforms

  • Renata F. MartinsEmail author
  • Marie-Louise Kampmann
  • Daniel W. Förster
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1963)

Abstract

Efficient methods for building genomic sequencing libraries from degraded DNA have been in place for Illumina sequencing platforms for some years now, but such methods are still lacking for other sequencing platforms. Here, we provide a protocol for building genomic libraries from degraded DNA (archival or ancient sample material) for sequencing on the Ion Torrent™ high-throughput sequencing platforms. In addition to a reduction in time and cost in comparison to commercial kits, this protocol removes purification steps prior to library amplification, an important consideration for work involving historical samples. Libraries prepared using this method are appropriate for either shotgun sequencing or enrichment-based downstream approaches.

Key words

Archival DNA Degraded samples Ion Torrent™ PGM Non-illumina platforms Sequencing libraries 

References

  1. 1.
    Pääbo S, Poinar H, Serre D et al (2004) Genetic analyses from ancient DNA. Annu Rev Genet 38(1):645–679CrossRefGoogle Scholar
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    Ion Torrent by Thermo Fischer (2016) Ion Xpress Plus gDNA fragment library preparation—user guide. Thermo Fischer. MAN0009847Google Scholar
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    Taylor PG (1996) Reproducibility of ancient DNA sequences from extinct Pleistocene fauna. Mol Biol Evol 13:283–285CrossRefGoogle Scholar
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    Maricic T, Whitten M, Pääbo S (2010) Multiplexed DNA sequence capture of mitochondrial genomes using PCR products. PLoS One 5(11):e14004CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Renata F. Martins
    • 1
    • 3
    Email author
  • Marie-Louise Kampmann
    • 1
    • 2
  • Daniel W. Förster
    • 1
  1. 1.Department of Evolutionary GeneticsLeibniz Institute for Zoo and Wildlife Research (IZW)BerlinGermany
  2. 2.Section of Forensic Genetics, Department of Forensic MedicineUniversity of CopenhagenCopenhagenDenmark
  3. 3.Institute for Biochemistry and BiologyUniversity of PotsdamPotsdamGermany

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