Tonight I watched an updated session entitled “How to get started with nanopore sequencing and plan your experiment.” The speaker was Akelia Wauchope-Odumbo, Associate Director of Technical Applications – Americas with Oxford Nanopore Technologies (ONT). Wauchope-Odumbo started this new masterclass series. The theme is “from sample to answer” covering preparation, sequencing, and analysis. The masterclass series will include a session on the NO-MISS bacterial genome sequencing workflow. Wauchope-Odumbo listed the objectives of this session, focusing on the use of ONT for simple projects. The main features of ONT sequencing were listed as direct sequencing of DNA and RNA, PCR-free, read-length agnostic, real-time analysis, and chemistry on bespoke electronics. The detection of base modifications is a key advantage of ONT. Wauchope-Odumbo noted that reads sequenced can span 20 bp to megabase reads. Wauchope-Odumbo provided suggestions for planning a sequencing experiment. They emphasized four steps, starting with the extraction of high-quality nucleic acids. Next, preparation of libraries for sequencing and sequencing. Finally, analysis of sequences. Wauchope-Odumbo emphasized that in most cases starting from the end is necessary. Planning your experiment should start with statement of the goals, sequences of interest, and size of the target/genome. Step one, preparing the input, takes into consideration the goals of the study and can utilize a library of protocols available on the Nanopore Community protocols website. For library preparation, ONT offers a series of kits for different applications. The data yield and number of runs needed will dictate the type of device needed. Wauchope-Odumbo shared a slide with unique environments in which researchers have successfully used ONT devices. For data analysis, ONT provides a range of data analysis options. The session ended with a recap of the numerous resources available on the ONT Community website. Every updated masterclass is full of new advice!
