Glowing Bacteria Lab

Part 1: Introduction

In this lab, we'll be performing what is known as "genetic transformation," which is the modification of genes in an organism. In the biotechnology industry, this is used in many applications, like genetically modified foods, bioremediation (using bacteria to digest oil spills,) and gene therapy in people with defective genes. We will be taking a Green Fluorescent Protein (GFP)-producing gene from glowing jellyfish, and inserting it into bacteria, which will incorporate the gene into its DNA, and produce the same glowing effect, and pass the trait on to its offspring.

 Part 2: Experiment

First we'll label both of our tubes: one as +pGLO and the other -pGLO, and our table number on both, then put them in the foam block. Then we'll put 250 microliters of transformation solution (calcium chloride) into each tube with a sterile pipet, then ice the tubes. Next we'll get a colony of bacteria from our starter plate with a sterile loop, and it put it in the +pGLO tube, then put the tube back on the ice. Then we'll repeat the process with the -pGLO tube, with a new sterile loop. Next we'll add 10 microliters of plasmid DNA to the +pGLO tube only, and flick the tube to mix it. Then we'll put the tubes on ice and let them sit for 10 minutes. While that's going on, we'll label our 4 agar plates. Next we'll do a heat shock treatment by taking the tubes from the ice to a 42 C waterbath for 50 seconds, then back onto the ice for 2 minutes. Then we'll add 250 microliters of LB broth to each tube, one at a time, with a sterile pipet. We'll then let the tubes sit at room temperature for 10 minutes. Next we'll mix the tubes by tapping them with a finger, and with a new sterile pipet for each tube, we'll pipet 100 microliters of the liquid onto the corresponding plates. Next, using a new sterile loop for each plate, we'll spread the liquid around the surface of the plate. Finally, we'll stack our plates, tape them together, put our table number on the bottom of the stack, and put the stack upside down in the 37 C incubator until Monday.


 Part 3: Results

Sure enough, the expected results of the lab were successful, and the bacteria glowed as expected. Since htis operation worked, I can't think of any possible margin of error, as there aren't any other stray glowing genes around that could have contaminated the sample. Our only slip-up was that the trays were left in the cold room all weekend, so the bacteria couldn't grow very quickly, and some probably died, but we did have some surviving glowing bacteria. This is probably my favorite lab we have done all year because of the level of technological advancement it uses-- it is the most current and relevant one that we have done in my opinion.