<aside> 💡 Improve Azotobacter's natural N fixation abilities by down-regulating nifL using CRISPR

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Overall, our experiment procedure goes as follows:

1. Order DNA constructs from Twist
2. Grow Azotobacter
3. Clone the cas9 and the gRNA together
4. Create CRISPR transformants and Cas9 transformants (control)
5. Test if the CRISPR is being expressed
6. Confirm the nifL knockout through sequencing
7. Assess the potential improvements through an ammonium assay

Protocols

1. Prepare culture medium
2. Culture Azotobacter cells
3. Create competent cells
4. Transform plasmid
5. Culture in antibiotic plates
6. Do assays

Inducing competency onto Azotobacter