Bacterial Isolate Genomic DNA Isolation – ONT NO-MISS

Your bacterial isolate has been grown on agar plates. We now need to lyse open the bacteria and isolate only the genomic DNA for sequencing. We need to lyse the bacteria with enzymes and a lysis buffer for this. We will then purify the genomic DNA. We will use a modification of the NEB Monarch Spin Column Kit extraction kit. This protocol has been recommended by Oxford Nanopore Technologies (ONT), the company that makes the sequencer we will use for sequencing our bacterial genomes.

On the left are healthy living bacteria (An example is Bacillus). On the right the bacteria are lysed open, which results in cell death. Consider this question as you read the protocol: How will we facilitate bacterial lysis? Images of Bacilli created using BioRender.com.

After completing this lab you will gain the following lab skills: