CRISPR Experiment
In this experiment we will use CRISPR to make a bacteria resistant to the antibiotic ampicillin. This is accomplished by replacing a part of DNA code that contains the directions for making the bacteria's ribosomes. It turns out that the bacteria's ribosomes are the target for ampicillin attacks. So we are going to change the design of the ribosomes by inserting a slightly different code into the bacterial DNA at the site where the directions for making the ribosome are located. That difference will make the bacterial ribosomes no longer vulnerable to Ampicillin.
Once we have made that change in the ribosome gene, we'll plant the transformed bacteria into a petri dish laced with Ampicillin. If the bacteria grow, then we know that our experiment was a success.

To begin this lab, start the program by clicking the link below and once it starts press the button

click here to start lab

1. Gloves on.
2. Set your pipette to 100 uL. and attach a pipette tip.
1. be sure your pipette is set to 100 uL and 2. only push the plunger to the first position (ie not all the way down) when you are drawing up transformation (PEG) from the original tube. Put this into an empty tube. (henceforth known as the transformation tube)


3. Here are the five steps involved in creating the transformation tube. Do one step at a time. Discard the pipette tip after every use in order to avoid cross contamination. Nota bene: DO NOT RETURN TO AN ORIGINAL TUBE WITH A PIPETTE TIP THAT HAS BEEN USED.


4. Now Put the tube of transformation mix into ice for 30 minutes.(in our case waiting that long will not affect the outcome, so set the timer for 30 minutes but after it runs a short time click the reset button to stop it)
5. When the 30 minutes is up, take the transformation tube from the ice and immerse it into luke warm water (108 deg F) for 30 seconds (set timer). Be sure the tube lid is on tight!!
6. Then, using a pipette transfer 100 ul from the tube of LB to the transformation tube we just removed from the warm water. We want to feed the little critters and let them relax after the trauma of the transformation.

7. Leave the tube to incubate all night at room temperature.

8. End of lab. press