37 Overview and Goals
Bacterial Genomic DNA Isolation – NEB HMW
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. For this, we need to gently lyse the bacteria with the enzyme lysozyme and then purify the genomic DNA. We will use the Monarch High Molecular Weight (HMW) DNA Extraction Kit (New England BioLabs) and protocol because we want long DNA strands to sequence with the Nanopore sequencer.
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:
Learning Objectives
- Lab safety and proper personal protective equipment (PPE)
- Proper use of a centrifuge and pipettors
- Isolation and purification of high-molecular weight genomic DNA
