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Emulsion PCR-coupled target enrichment: an effective fishing method for high-throughput sequencing of poorly preserved ancient DNA.

Authors:
Makio Kihana Fuzuki Mizuno Rikai Sawafuji Li Wang Shintaroh Ueda

Gene 2013 Oct 27;528(2):347-51. Epub 2013 Jul 27.

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.

Due to the difficulties in deep sequencing, high-throughput sequencing of ancient DNA has been limited to exceptionally well-preserved ancient materials. The primary factor is microbial attack popularly observed in the buried materials, and it causes drastic increase in relative ratio of microbial DNA in the extracted DNA. We present a unified strategy in which emulsion PCR is coupled with target enrichment followed by next-generation sequencing. The method made it possible to obtain efficiently non-duplicated reads mapped to target sequences of interest, and this can achieve deep and reliable sequencing of ancient DNA from typical materials, even though poorly preserved.

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http://dx.doi.org/10.1016/j.gene.2013.07.040DOI Listing
October 2013

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