Daca-Roszak Patrycja, Fiedorowicz Joanna, Jankowski Maciej, Ciesielka Marzanna, Teresiński Grzegorz, Lipska-Zietkiewicz Beata, Zietkiewicz Ewa, Grzybowski Tomasz, Skonieczna Katarzyna
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.
Department of Animal Physiology, Biochemistry and Biostructure, Poznan University of Life Science, Poznan, Poland.
J Appl Genet. 2024 Sep;65(3):559-563. doi: 10.1007/s13353-023-00821-4. Epub 2023 Dec 19.
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.
大规模平行测序（MPS）技术因其在检测线粒体DNA异质性方面的高灵敏度，已成为线粒体DNA研究的金标准，线粒体DNA异质性是各种医学应用中的一种预后标志物。存在多种用于线粒体DNA分析的MPS技术和平台。要获得可靠且灵敏的结果，需要对整个线粒体DNA序列进行深度且均匀的覆盖，而这在很大程度上受到文库制备方法和测序平台选择的影响。在此，我们比较了使用两种文库制备方案（Nextera XT DNA文库制备试剂盒和Nextera DNA Flex文库制备试剂盒）以及两种不同的Illumina MPS平台（MiSeq FGx和ISeq 100）时的测序覆盖度和异质性检测能力。我们的研究表明，Nextera DNA Flex文库方案在整个线粒体基因组上提供了更平衡的覆盖度，并且在MiSeq和ISeq Illumina MPS系统中均能实现可靠的异质性检测。
