Pglo Lab Report

The pGLO lab is a lab where students attempt to put the genes that make a jelly fish glow into E. Coli. After a process called transformation, the process in which a cell takes up and expresses a new piece of genetic information, the E. Coli will be able to glow and will be antibiotic resistant. The students first need to learn a couple of techniques before they are able to begin this lab. The first technique they will need is how to keep their environment sterile. They must learn to only open their tools, and other supplies for as long as they need, and keep them from touching anything. They must also learn that tools can never be shared between specimens, and when they use a tool, they should dispose of it and get a new one. Next they need to learn how to transfer a bacterial colony. To do this, one must use a sterile loop and scrape one colony off of the dish. Then they must insert the loop into
Then we opened the tubes and using a sterile pipet we put 250 µl of transfer solution in and placed them on ice. Next we removed them from the ice and used a sterile loop to pick up a single colony of bacteria. We put a colony in both tubes and then placed both tubes back on the ice. After that, we placed a loopful of plasmid DNA into the positive pGLO. We then incubated the tubes on ice for ten minutes. After the ten minutes were up, we placed the tubes in a bath of forty two degree centigrade water for fifty seconds, and then quickly back onto the ice for two minutes. After that we removed them from the ice and added 250 µl of LB nutrient broth to the tubes and let them sit at room temperature for ten minutes. When the ten minutes had passed, we flicked the tubes to mix them and added 100 µl of transformation and control suspensions onto the appropriate plates. Finally we spread the solution using a sterile loop, stacked the plates, and placed them upside down in an incubator at thirty seven degrees