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A method for single-stranded ancient DNA library preparation

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Gansauge,  Marie-Theres       
Advanced DNA Sequencing Techniques, Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Meyer,  Matthias
Advanced DNA Sequencing Techniques, Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Citation

Gansauge, M.-T., & Meyer, M. (2019). A method for single-stranded ancient DNA library preparation. In B. Shapiro, A. Barlow, P. D. Heintzman, M. Hofreiter, J. L. A. Paijmans, & A. E. R. Soares (Eds.), Ancient DNA: Methods and Protocols (2nd ed., pp. 75-83). New York, NY: Springer.


Cite as: https://hdl.handle.net/21.11116/0000-0003-4B3D-6
Abstract
Genomic library preparation from highly degraded DNA is more efficient when library molecules are prepared separately from the complementary strands of DNA fragments. We describe a protocol in which libraries are constructed from single DNA strands in a three-step procedure: single-stranded ligation of the first adapter with T4 DNA ligase in the presence of a splinter oligonucleotide, copying of the DNA strand with a proofreading polymerase, and blunt-end ligation of the second double-stranded adapter with T4 DNA ligase.