Extraction and Oxford Nanopore sequencing of genomic DNA from filamentous Actinobacteria

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...

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Detalles Bibliográficos
Autores principales: Alvarez-Arevalo, Maria, Sterndorff, Eva Baggesgaard, Faurdal, David, Jørgensen, Tue Sparholt, Mourched, Anna-Sophie, Vuksanovic, Oliwia, Saha, Subhasish, Weber, Tilmann
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9792953/
https://www.ncbi.nlm.nih.gov/pubmed/36527715
http://dx.doi.org/10.1016/j.xpro.2022.101955
Descripción
Sumario: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.

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