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

dc.abstract.enMassively 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 infuenced 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 diferent (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
dc.affiliationWydział Medycyny Weterynaryjnej i Nauk o Zwierzętach
dc.affiliation.instituteKatedra Fizjologii, Biochemii i Biostruktury Zwierząt
dc.contributor.authorDaca-Roszak, Patrycja
dc.contributor.authorFiedorowicz, Joanna
dc.contributor.authorJankowski, Maciej
dc.contributor.authorCiesielka, Marzanna
dc.contributor.authorTeresiński, Grzegorz
dc.contributor.authorLipska-Zietkiewicz, Beata
dc.contributor.authorZietkiewicz, Ewa
dc.contributor.authorGrzybowski, Tomasz
dc.contributor.authorSkonieczna, Katarzyna
dc.date.access2024-10-03
dc.date.accessioned2024-10-03T09:53:09Z
dc.date.available2024-10-03T09:53:09Z
dc.date.copyright2023-12-19
dc.date.issued2024
dc.description.abstract<jats:title>Abstract</jats:title><jats:p>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 (<jats:italic>Nextera XT DNA Library Preparation Kit</jats:italic> and <jats:italic>Nextera DNA Flex Library Preparation Kit</jats:italic>) 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.</jats:p>
dc.description.bibliographyil., bibliogr.
dc.description.financepublication_nocost
dc.description.financecost0.00
dc.description.if2,0
dc.description.points140
dc.description.reviewreview
dc.description.versionfinal_published
dc.description.volume65
dc.identifier.doi10.1007/s13353-023-00821-4
dc.identifier.eissn2190-3883
dc.identifier.issn1234-1983
dc.identifier.urihttps://sciencerep.up.poznan.pl/handle/item/1767
dc.identifier.weblinkhttps://link.springer.com/article/10.1007/s13353-023-00821-4
dc.languageen
dc.relation.ispartofJournal of Applied Genetics
dc.relation.pages559-563
dc.rightsCC-BY
dc.sciencecloudnosend
dc.subject.enmtDNA
dc.subject.enHeteroplasmy detection
dc.subject.enMPS library preparation
dc.subject.enNextera Flex
dc.subject.enNextera XT
dc.titleThe effect of library preparation protocol on the efficiency of heteroplasmy detection in mitochondrial DNA using two massively parallel sequencing Illumina systems
dc.typeJournalArticle
dspace.entity.typePublication
oaire.citation.issue3
oaire.citation.volume65