New insights on single-stranded versus double-stranded DNA library preparation for ancient DNA. (December 2015)

• Record Type: Journal Article
• Authors: Wales, Nathan; Carøe, Christian; Sandoval-Velasco, Marcela; Gamba, Cristina; Barnett, Ross; Samaniego, José Alfredo; Ramos Madrigal, Jazmín; Orlando, Ludovic; Gilbert, M. Thomas P.
• Is Part Of: Biotechniques. Volume 59:Number 6(2015)
• Page Start: 368
• Page End: 371
• Publication Date: 2015-12
• Subjects: ancient DNA -- paleogenomics -- DNA library preparation -- next-generation sequencing; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000114364
• Languages: English
• ISSNs: 0736-6205

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.