Abstract
A detailed protocol is presented for the targeted enrichment of whole mitochondrial genomes based on an in-solution hybridization strategy. Bait is produced in-house by sonication of two long-range PCR amplicons and ligation of biotinylated double-stranded adapters. Indexed target DNA is hybridized with the bait in a multiplex enrichment reaction and pulled down using magnetic streptavidin beads followed by subsequent post-enrichment PCR and sequencing on an Illumina MiSeq. This strategy removes the need for expensive commercial bait probes while allowing enrichment of multiple samples in a single hybridization reaction. The method is particularly suitable for degraded DNA as it is able to enrich short DNA fragments and is not susceptible to polymerase artifacts introduced during PCR-based assays.
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Bekaert, B., Ellerington, R., Van den Abbeele, L., Decorte, R. (2016). In-Solution Hybridization for the Targeted Enrichment of the Whole Mitochondrial Genome. In: Goodwin, W. (eds) Forensic DNA Typing Protocols. Methods in Molecular Biology, vol 1420. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3597-0_14
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DOI: https://doi.org/10.1007/978-1-4939-3597-0_14
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3595-6
Online ISBN: 978-1-4939-3597-0
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