So last week I began work on the Crispr kit provided by the Odin, intended to teach Crispr by having the user genetically engineer bacteria to produce GFP. For those who don’t know, GFP stands for green fluorescent protein, which means it will appear to glow green when exposed to UV light.
The initial step of the project involves creating agar plates and adding a laboratory (safe) strain of e.coli to them. Here are the initial plates.
The instructions provided by the Odin involve measuring out a specific amount of LB-Agar mix with 150mL of water and microwaving them until the mix turns a clear yellow-brownish color. The amount of mix provided with the kit is not that much, so when you want to learn to grow your own yeast and bacteria, you’ll want to check out the sources below.
Anyway, I’ve been holding onto this kit for a year without touching it, and bacteria stabs may not preserve well for that long. Particularly as at one point the bag (even though it said DO NOT FREEZE on it was mistaken for the one that belonged in the freezer, so these bacteria were frozen for some months.
The kit included two different vials of E.Coli, plus a bacteria called PDawn and PDusk (to my knowledge these were supposed to be pre-engineered versions so that you would have something to compare yours to). After plating those, three agar plates remained.
I remembered an experiment done with children demonstrating just how dirty our hands are, so I stuck my thumb in one plate, spat in another and left the final one blank to act as a control (after all, my apartment is far from sterile).
After a few days growth was visible on the PDawn, PDusk, spit and thumb plates, but no apparent growth on the E.Coli plates. It his been a week since then. Here is the current condition.
Since no apparent growth has taken place for the E.Coli (what you see in the corner in the plate is damage from when I dropped it), the Crispr experiment will have to be postponed until I get it fresh (that sounds so weird saying it out loud). With the remainder of our time, let’s look at these samples under a microscope and see if we can discover anything interesting about the bacteria.
The first thing i noticed was that the plates had a strong smell to them. Agar itself is a smelly broth, hard to describe as anything except bland yet unappetizing, adjectives which also applied to all of the bacterial growth colonies. Surprise surprise, the one I spat in still smelled faintly of spit, and the one labelled PDusk smelled like feet.
Next are some pictures under a microscope to see how they look. First we’ll look at PDawn, since it kinda looks the most grainy.
The discolorations are the closest you can see of bacterial growth. It’s a little more evident in the PDusk variety.
Next is the spit.
Saving the best for last, we have the colonies produced by my thumb. There I can discern at least three different ones. The yellow, the whitish and the ones that seem to make wider glops.
Here are the attached images at varying magnifications. Let’s see if you can guess which is which.
In a week or two I hope to give you an update on the Crispr experiment, but if not, then as a consolation prize I’ll definitely get pictures of more cool things to see under a microscope. With luck, maybe they won’t smell like feet!
You can find the Odin’s Crispr kit here: https://www.the-odin.com/diy-crispr-kit/
This is where I will be purchasing fresh E.Coli: https://www.homesciencetools.com/product/escherichia-coli-bacteria/