The Experiment: Genetically Modifying Bacteria

This is a part 2 of my experiment with genetically modifying E.Coli to be bioluminescent and express the GFP from jellyfish. If you have yet to check part 1 out I would definitely recommend doing so, it explains the science behind what we’re doing and why it’s important.

These were the bacteria after my experiment didnt work…

Let’s talk about how the experiment went for me!

All in all, it simply didn’t work — as I was expecting and had heard from other people who had also tried it. It was a great experience for me to have a test of my own and troubleshoot, but I didn’t get any clear-cut results or cool glowing bacteria.

The experiment essentially *should* have gone as follows:

1. Agar with and without ampicillin was prepared
2. The bacteria was ‘planted’ on the agar and in 1–2 days, growth occurred in which colonies formed
3. Bacteria was scraped off of the agar and the plasmid was activated
4. The competent cell mixture was incubated
5. An LB powder micro-centrifuge bottle was added into the bottle and it was left to incubate
6. This was then put onto the LB Kan Agar plate (which contains the ampicillin that the bacteria that absorbed the plasmids would have resistance to)
7. It was incubated for 2ish days and then when observed with the blue light, it glowed fluorescent green

The ODIN instructions

Instead, it went like this:

Steps 1–6 were completed, and after 2 days when observed with the blue light, there was no fluorescent colouring.

I reached out to the Odin and have yet to hear a response. I cant say it’s a surprise as the people who I spoke with who had already completed the experiment had similar results (I learned this after I purchased it).

I had very minimal bacterial growth even with using multiple test plates to try and redoing the agar twice. After I had bacterial growth and followed all of the other steps correctly, I still was left with bacteria that were unable to glow, which makes me think there was either something wrong with the bacteria themselves (in that they were not frozen during transportation), or something wrong with the way I incubated the bacteria?

I will be trying the experiment again after consulting more employees at the Odin and reviewing the steps I took — hopefully this article will be replaced with one that has photos of glowing bacteria (that are actually mine!)

Thanks for reading; if you’ve tried this experiment let me know how it went for you! Feel free to check out my other articles and have a wonderful day!