We invite you to tour Next Generation Sequencing (NGS) from Oxford Nanopore Technologies (ONT). As one of the most captivating newly emerged practices in genomics research, ONT sequencing grants the researchers real-time DNA and RNA analysis with accuracy and convenience.

If you are considering integrating ONT sequencing into work in your lab, here are some practical first steps:

  • Learn the Basics: Before proceeding with ONT sequencing, comprehend what underpins the technology. Understand nanopore sequencing technology, its products, and workflows and review the available resources such as webinars, tutorials, and user manuals.
  • Establish Your Research Needs: Determine how ONT sequencing can fit into the research goals of your lab. Whether you are interested in whole-genome sequencing, targeted sequencing, RNA sequencing, or epigenetics analysis, ONT provides many different kits and protocols that will work for almost any application.
  • Invest in the Right Equipment: To perform ONT sequencing, you’ll need the necessary equipment, including a nanopore sequencer (e.g., MinION, GridION, or PromethION), a computing device for data analysis, and associated accessories such as flow cells and reagents. Consider your requirements and budget constraints when selecting the appropriate sequencer model.

Sample Preparation in Optimal Manner

Sample preparation is critical to success in ONT sequencing. Follow the recommended sample preparation protocols for DNA or RNA extraction, library construction, and sequencing library loading, paying close attention to factors like sample quality, quantity, and purity toward optimal sequencing results.

Quality Control

Carefully check the quality of your samples and sequencing libraries before sequencing. Evaluate the integrity of DNA or RNA through electrophoresis methods in agarose gel, qPCR, or fluorometric analysis. Check library concentration and size distribution to ensure even sequencing coverage.

Sequence Wisely

Design your sequencing experiments to get the actual numbers needed at the required sequencing depth with minimum experimental noise. For this purpose, the sequencing depth, read length, and sequencing approach (i.e., 1D or 1D^2) are considered based on the study requirements and sample-related parameters.

Keep Up With Software Tools

Get the latest software tools and analysis pipelines for ONT sequencing data handling. Explore what platforms will be helpful for alignment, variant calling, and other bioinformatics analyses, like the EPI2ME platform, Guppy base-calling software, and Nanopack processing toolkit. Also, use community-supported tools and resources that target specific applications or research domains.

Validity Testing

Confirm your sequencing results through independent means such as PCR, Sanger sequencing, or alternative sequencing methods, including Illumina or PacBio. Cross-validate your ONT data with publicly available datasets or previously published results to confirm the accuracy and reproducibility of your results.

Troubleshoot Effectively

Be prepared to troubleshoot potential technical issues while running your samples. Be prepared and know the standard troubleshooting practices such as priming flow cells, cleaning pores, and normalizing signals when sequencing on nanopore. If you have questions, refer to ONT’s support documentation or contact the user community.

Be patient and persistent

As with any other technology, using ONT sequencing takes time to master and much practice, not derailed by early difficulties or forced errors; get feedback on learning from others who have gone before you.

Follow these practical tips and be systematic in your actions. You’ll accomplish integrating ONT sequencing with the rest of the protocols in your laboratory, leading to new insights into genome and transcriptome complexity. Happy sequencing!