Dec 15, 2015

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

Nathan WalesM Thomas P Gilbert


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.

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  • Citations6


Mentioned in this Paper

AdnA protein, Pseudomonas fluorescens
Research Personnel
Ancient DNA
Sequence Determinations, DNA
probe gene fragment
cDNA Library
Nucleic Acid Strand
Bowel Preparation
DNA, Single-Stranded

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