Chapter title |
In-Solution Hybridization for the Targeted Enrichment of the Whole Mitochondrial Genome.
|
---|---|
Chapter number | 14 |
Book title |
Forensic DNA Typing Protocols
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3597-0_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3595-6, 978-1-4939-3597-0
|
Authors |
B. Bekaert, R. Ellerington, L. Van den Abbeele, R. Decorte |
Editors |
William Goodwin |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Portugal | 1 | 10% |
Unknown | 9 | 90% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 3 | 30% |
Librarian | 1 | 10% |
Professor | 1 | 10% |
Other | 1 | 10% |
Student > Ph. D. Student | 1 | 10% |
Other | 1 | 10% |
Unknown | 2 | 20% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 20% |
Medicine and Dentistry | 2 | 20% |
Immunology and Microbiology | 1 | 10% |
Agricultural and Biological Sciences | 1 | 10% |
Unknown | 4 | 40% |