Extraction and Oxford Nanopore sequencing of genomic DNA from filamentous Actinobacteria

Maria Alvarez-Arevalo, Eva Baggesgaard Sterndorff, David Faurdal, Tue Sparholt Jørgensen*, Anna-Sophie Mourched, Oliwia Vuksanovic, Subhasish Saha, Tilmann Weber*

*Corresponding author for this work

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Actinomycetota (Actinobacteria) is an ecologically and industrially important phylum which is challenging to extract pure high-molecular-weight (HMW) DNA from. This protocol provides a parallelized, cost-effective, and straightforward approach for consistently extracting pure HMW DNA using modified non-toxic commercial kits suitable for higher throughput applications. We further provide a workflow for sequencing and assembly of complete genomes using an optimized Oxford Nanopore rapid barcoding protocol and Illumina data error correction.
Original languageEnglish
Article number101955
JournalSTAR Protocols
Issue number1
Number of pages24
Publication statusPublished - 2023


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