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

作者：Wales Nathan^*; Caroe Christian; Sandoval Velasco Marcela; Gamba Cristina; Barnett Ross; Samaniego Jose Alfredo; Madrigal Jazmin Ramos; Orlando Ludovic; Gilbert Thomas P

来源：Biotechniques, 2015, 59(6): 368-371.

DOI：10.2144/000114364

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

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.

• 出版日期2015-12