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Biotechniques. 2015 Dec 1;59(6):368-71. doi: 10.2144/000114364. eCollection 2015 Dec.

New insights on single-stranded versus double-stranded DNA library preparation for ancient DNA.

Author information

1
Center for GeoGenetics, Natural History Museum of Denmark, University of Copenhagen, Copenhagen, Denmark.
2
Center for Biological Sequence Analysis, Technical University of Denmark, Kgs. Lyngby, Denmark.
3
Trace and Environmental DNA Laboratory, Department of Environment and Agriculture, Curtin University, Perth, Australia.

Abstract

An innovative single-stranded DNA (ssDNA) library preparation method has sparked great interest among ancient DNA (aDNA) researchers, especially after reports of endogenous DNA content increases >20-fold in some samples. To investigate the behavior of this method, we generated ssDNA and conventional double-stranded DNA (dsDNA) libraries from 23 ancient and historic plant and animal specimens. We found ssDNA library preparation substantially increased endogenous content when dsDNA libraries contained <3% endogenous DNA, but this enrichment is less pronounced when dsDNA preparations successfully recover short endogenous DNA fragments (mean size < 70 bp). Our findings can help researchers determine when to utilize the time- and resource-intensive ssDNA library preparation method.

KEYWORDS:

DNA library preparation; ancient DNA; next-generation sequencing; paleogenomics

PMID:
26651516
DOI:
10.2144/000114364
[Indexed for MEDLINE]
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