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

Nathan Wales | Zendy; Christian Car⊘e | Zendy; Marcela SandovalVelasco | Zendy; Cristina Gamba | Zendy; Ross Barnett | Zendy; José Alfredo Samaniego Castruita | Zendy; Jazmín RamosMadrigal | Zendy; Ludovic Orlando | Zendy; M. Thomas P. Gilbert | Zendy

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

Author(s) -

Nathan Wales,

Christian Car⊘e,

Marcela SandovalVelasco,

Cristina Gamba,

Ross Barnett,

José Alfredo Samaniego Castruita,

Jazmín RamosMadrigal,

Ludovic Orlando,

M. Thomas P. Gilbert

Publication year - 2015

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/000114364

Subject(s) - dna , endogeny , double stranded , ancient dna , biology , library , genomic library , microbiology and biotechnology , dna synthesis , computational biology , genetics , biochemistry , gene , base sequence , population , 16s ribosomal rna , demography , sociology

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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