Comparison of different methods to isolate high-molecular-weight DNA from bacteria for nanopore sequencing

Nanopore long-read sequencing has overcome many of the restrictions of short-read sequencing, enabling the resolution of repetitive regions and the assembly of closed bacterial genomes.

This has allowed more researchers to make use of long-read technologies for a variety of applications. Besides the careful handling of samples in library preparation, the most important prerequisite to achieve optimal sequencing results is the quality and size of the input DNA. Size distribution has a major impact on the read length and therefore the quality of the assembly and success of resolving large repetitive regions. The proper isolation of high-quality DNA of optimal size is therefore crucial for successful Nanopore sequencing.

The German NGS Competence Center in Tübingen tested different DNA isolation kits from different manufacturers mostly dedicated for high-molecular weight DNA isolation and performed quality control analysis using the Agilent Femto Pulse system to determine the size and distribution of the isolated genomic DNA.

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