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Cost-effective straightforward method for captured whole mitogenome sequencing of ancient DNA
A. Senovska, E. Drozdova, O. Vaculik, F. Pardy, K. Brzobohata, D. Fialova, J. Smerda, P. Kos
Jazyk angličtina Země Irsko
Typ dokumentu časopisecké články
NLK
ProQuest Central
od 1997-02-07 do Před 2 měsíci
Nursing & Allied Health Database (ProQuest)
od 1997-02-07 do Před 2 měsíci
Health & Medicine (ProQuest)
od 1997-02-07 do Před 2 měsíci
- MeSH
- analýza nákladů a výnosů MeSH
- genom mitochondriální * MeSH
- lidé MeSH
- mitochondriální DNA genetika MeSH
- polymerázová řetězová reakce MeSH
- soudní genetika metody MeSH
- starobylá DNA * MeSH
- vysoce účinné nukleotidové sekvenování metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Working with mitochondrial DNA from highly degraded samples is challenging. We present a whole mitogenome Illumina-based sequencing method suitable for highly degraded samples. The method makes use of double-stranded library preparation with hybridization-based target enrichment. The aim of the study was to implement a new user-friendly method for analysing many ancient DNA samples at low cost. The method combines the Swift 2S™ Turbo library preparation kit and xGen® panel for mitogenome enrichment. Swift allows to use low input of aDNA and own adapters and primers, handles inhibitors well, and has only two purification steps. xGen is straightforward to use and is able to leverage already pooled libraries. Given the ancient DNA is more challenging to work with, the protocol was developed with several improvements, especially multiplying DNA input in case of low concentration DNA extractions followed by AMPure® beads size selection and real-time pre-capture PCR monitoring in order to avoid cycle-optimization step. Nine out of eleven analysed samples successfully retrieved mitogenomes. Hence, our method provides an effective analysis of whole mtDNA, and has proven to be fast, cost-effective, straightforward, with utilisation in population-wide research of burial sites.
CF Genomics CEITEC Masaryk University Kamenice 753 5 Brno 62500 Czech Republic
Institute for Archaeological Heritage Kaloudova 1321 30 Brno 61400 Czech Republic
Masaryk Museum Zamecke Nam 27 9 Hodonin 69501 Czech Republic
Citace poskytuje Crossref.org
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