- Brett Fuller
- Holly Tjaden
- Chase Meusel
Tetrodotoxin Production in E. coli Using Flp Genes from Pufferfish
- 1. Obtain live puffer fish specimen and extract the total genomic DNA from fin tissue.
- 2. Design primers to amplify the three flp genes identified as well as a gene for orange fluorescence to confirm that the flp genes should be getting expressed. These primers should also contain the restriction site ends that are required for biobrick usage.
- 3. Amplify all three flp genes using PCR from the total genomic DNA isolated from the puffer fish tissue.
- 4. Amplify the orange fluorescence gene from the biobrick plasmid using the provided procedure.
- 5. Ligate the orange fluorescence gene and one or more of the flp genes.
- 6. Ligate the combined genes into a plasmid and transform the plasmid into the bacteria.
- 7. Allow the bacteria to grow up and plate them on selective media.
- 8. Test bacterial colonies for fluorescence using UV radiation.
Important Results and Milestones
- keep track of your most important results and refer to the corresponding page in your notebook
- upload important pictures (don't forget to label them! Powerpoint is very convenient). Remember: these will become quite handy later in your summary report or final presentation. If you do label and upload the pictures as soon as you got them, your summary report can be written much more effortlessly (do you usually procrastinate? This is chance to do some work before hand that frees you up for finals week).
Introduction Powerpoint to Project
Final Powerpoint Presentation