Pglo Lab Report

Transformations Presented in E. Coli Using the pGlo Gene
Introduction
In biology, genetic transformations involve the transfer of a gene into an organism to give that organism a new gene to express (Blaber, 2004). Genetic transformations can be used for therapy, vaccinations, and in tissue renewal (Rivera,2014). In 1928, a man named Frederick Griffith discovered the term genetic transformations and their use (Griffiths,2000). To test these new genetic transformations a substance called pGlo is used to allow for replication and to see the results because of the bioluminescence. From Genetic Transformation, Dr. Michael Blaber wrote, “The pGLO plasmid contains an origin or replication, a selectable marker, and the gene for Green Fluorescent

As a result of the pGlo gene being inserted into the nonpathogenic substance of E.Coli, this experiment is testing whether or not E.Coli will grow and be bioluminescent. The independent variable in this experiment would be the E.Coli because it is either going to reproduce or not reproduce. Also, the pGlo gene is an independent variable because as a result the E.Coli is either going to be glowing or not glowing. The dependent variable would be the number of bacterial growth with the 12-18 hour time

One needs to be labeled +PGlo and the other container is labeled -Pglo. Next, in both test tubes using a dropping, 250 microliters of calcium chloride is needed (Making sure to use a new dropper after each test tube). Then place both containers into a styrofoam cup containing crushed ice a. Taking a sterile loop, scrap a thin layer of E.coli from the petri dish onto the loop (making sure the loop gets a clear film over it). Remove the +pGlo test tube and open the container. Place the loop with the nonpathogenic E.coli and twirl it around in the test tube. Next, remove the loop and close container, place the +pGlo container back into the cup full of ice. Next, remove the test tube labeled -pGlo and repeat the instructions listed above making sure to get a new sterile loop before continuing. Then place the -pGlo test tube container back onto the ice and examine the pGlo genes. Using the pGlo gene, put one loop full of the pGlo into the container labeled +pGlo (Make sure to use a sterile loop). Place back on the crushed ice and wait ten minutes. While waiting, label the four petri dishes B lactamase - pGlo, B lactamase/ ampicillin -pGlo, B lactamase / ampicillin +pGlo ,and B lactamase/arabinase/ ampicillin +pGlo. After labeling the petri dishes, remove -pGlo and +pglo test tube containers from the ice cup and place them on the heat rack (at 42℃). Only leave the test tubes on for 50 seconds. After