CRISPR LAB!

Emilia Syvertsen
Dec 30, 2023
2 min read

Updated: Jun 10, 2024

Hi! This post is going to be about...my crispr lab.

So first there are 4 stages ; 1 week before, 3 days before, 1 day before, and the day of the lab.The point of the experiment is to break the DNA then substitute it to make the fungus blue, instead of white.This is a picture of what it should look like when it is done:

A. MAKE PLATES

These are the observations for day one:

It is pretty easy to do so don't sweat it

warm up the distilled water and add LB agar powder. Swirl it around. Microwave in 30-second increments just until it boils.

Next add the ChIX (which I call CheX mix) then swirl again. Do that a couple more times. Add LB agar soloution. Pour into the petri dishes.

B. REHYDRATE

BACTERIA

Uncap the vial of lyophilized bacteria and add LB + ampiccillin close the vial. Shake it softly by flicking it. Add 50 μl of rehydrated bacteria into 16 sterile 1.7 ml microtubes. Incubate at room temperature for 48-60 hours.

Do not disturb or agitate samples during the incubation period.

C. ADD CaCI2

TO BACTERIA

Add 25 μl of sterile CaCl2 to each tube of bacteria. Place tubes in refrigerator and chill for 12-24 hours. (Best results are observed after a 12 hour incubation).

D. PREP SUPPLIES

& EQUIPMENT

So basically this is what happens:

The DNA is sitting there and a certain part of it is the LacZ gene and a RNA comes along and copes the LacZ gene and pastes it into the ribazomes to create blue proteins. In this experiment we add Cas9 with its guide RNA that comes along and finds the LacZ gene and cuts it, and it turns white insteade of blue because it cut the LacZ gene that codes for the blue proteins.