The effect of library preparation protocol on the efficiency of heteroplasmy detection in mitochondrial DNA using two massively parallel sequencing Illumina systems.

Autor: Daca-Roszak P; Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland., Fiedorowicz J; Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.; Department of Animal Physiology, Biochemistry and Biostructure, Poznan University of Life Science, Poznan, Poland., Jankowski M; Department of Biology and Medical Genetics, Medical University of Gdansk, Gdansk, Poland., Ciesielka M; Chair and Department of Forensic Medicine, Medical University of Lublin, Lublin, Poland., Teresiński G; Chair and Department of Forensic Medicine, Medical University of Lublin, Lublin, Poland., Lipska-Zietkiewicz B; Centre for Rare Diseases, Medical University of Gdansk, Gdansk, Poland; Clinical Genetics Unit, Department of Biology and Medical Genetics, Medical University of Gdansk, Gdansk, Poland., Zietkiewicz E; Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland., Grzybowski T; Department of Forensic Medicine, Faculty of Medicine, Collegium Medicum, Nicolaus Copernicus University, Bydgoszcz, Poland., Skonieczna K; Department of Forensic Medicine, Faculty of Medicine, Collegium Medicum, Nicolaus Copernicus University, Bydgoszcz, Poland. katarzyna.skonieczna@cm.umk.pl.
Jazyk: angličtina
Zdroj: Journal of applied genetics [J Appl Genet] 2024 Sep; Vol. 65 (3), pp. 559-563. Date of Electronic Publication: 2023 Dec 19.
DOI: 10.1007/s13353-023-00821-4
Abstrakt: Massively parallel sequencing (MPS) technology has become the gold standard in mitochondrial DNA research due to its high sensitivity in detecting mtDNA heteroplasmy, a prognostic marker in various medical applications. Various MPS technologies and platforms used for mtDNA analysis exist. Obtaining reliable and sensitive results requires deep and uniform coverage of the entire mtDNA sequence, which is heavily influenced by the choice of library preparation method and sequencing platform. Here, we present a comparison of the sequencing coverage and the ability to heteroplasmy detection using two library preparation protocols (Nextera XT DNA Library Preparation Kit and Nextera DNA Flex Library Preparation Kit) and two different (MiSeq FGx and ISeq 100) Illumina MPS platforms. Our study indicates that the Nextera DNA Flex Library protocol provides a more balanced coverage along the mitogenome and a reliable heteroplasmy detection with both MiSeq and iSeq Illumina MPS systems.
(© 2023. The Author(s).)
Databáze: MEDLINE
Externí odkaz: