54 Overview and Goals
Rapid Barcode Sequencing with Nanopore MinION
You have isolated, purified, and quantified high-molecular weight DNA from your bacterial isolate. You have also worked with your bacterium and grown in the presence of different carbon sources and pH levels to characterize its properties using Biolog GEN3 plates. Now, we have the opportunity to sequence the DNA of your isolate using long-read sequencing and Oxford Nanopore Technologies (ONT) MinION. With the genetic sequences we obtain, we will attempt to assemble the genome of this organism and explore its genomic potential!
The MinION sequencer from Oxford Nanopore Technologies (ONT) will allow us to sequence the genomic DNA using a flow cell with hundreds of nanopores. The DNA will be passed through the pores, and changes in current will be interpreted as bases (“base calling”). After performing quality control to remove low quality sequences, we will assemble the sequences to create contigs and scaffolds with the goal of exploring the genome of your isolates. Image created with BioRender.com.
After completing this lab you will gain the following lab skills:
Learning Objectives
- Lab safety and proper personal protective equipment (PPE)
- Pipetting with micropipettes
- Preparing sequencing libraries for the ONT MinION sequencing device
