Glowing Bacteria: The Future is Bright

Genetic transformation is a process in which one can change an organsim through the genes. Scientists recently began experimenting with this process to give new characteristics to living organisms by combining genes. By cutting, circular strands of DNA called plasmids with restriction enzymes, certain genes can be taken out and then combined with other cut plamids. DNA ligase is then added to seal the new strand and the new plasmid is completed. Scientists have many applications for this technique such as gentically engineering plants to be resistant to frost, insects, disease, etc. In this lab, we will first use a transfer pipet to transfer 250 µl of transformation solution (CaCl₂) into two tubes, one labeled +pGLO and the other labeled
-pGLO.  Then, after placing the tubes on ice, we will use a sterile loop to pick up bacteria from our starter plate and submerge them completely in each tube (using a different loop for each). Then we will ad 10 µl of plasmid DNA directly to liquid into just the +pGLO tube and flick the tube gently yo mix. Next we will return both the tubes to the ice for 10 minutes. After labeling four agar plates, we will begin to heat shock the tubes. By moving them from ice to a 42 degree celcius water bath, the phospholipid bilayer of the bacteria will stop repelling the plasmids and they will be able to enter the bacteria.  Then we will add 250 µl of LB nutrient broth to each tube. Finally, we will use new pipets to transfer 100 µl of each tube onto the appropriate plates and spread it around. The normal, untransformed bacteria should be killed by the ampicillin but the transformed bacteria has a resistance to this antibiotic. Therefore, this plate will glow a fluorescent green under UV light.