E. Coli Experiment

This experiment tested the growth of E.coli with inserted plasmid on an agar plate with Ampicillin. One colony of E.coli resistant to Ampicillin was grown during this experiment. The overall goal of the experiment was to successfully grow E.coli on the agar plate, which would show that the plasmid had been effectively inserted into the bacteria’s genes. This experiment helped students understand how plasmids were inserted into bacteria and used in real life situations. It also showed how the bacteria’s genes can be manipulated and changed to have new traits not usually present in bacteria. The students were able to compare regular E.coli with the E.coli they inserted plasmids into, which gave a better look into how the genes of the bacteria
The instruments and tools used may not have been completely sterile or cleaned, which could result in skewed data. Some procedural errors such as, time restrictions in the ice bath treatment and not heating the E.coli long enough, could have had some effect on the rate of recombination or the presence of recombination overall. The measurements used in this experiment, were not completely accurate, which most likely also led to error. Although, there was a small amount of resistant E.coli grown, there must have been some errors that took place because of the considerably small amount
A sterile syringe was then used to add 0.25 mL of calcium chloride to both tubes. The starter plate needed contained the colony of E.coli needed for the experiment. A sterile innoculating loop was gently scraped over the colony and transferred into the tube with the calcium chloride, by gently twirling the tube in the solution. Using the capillary micropipette and plunger, 10 picometers of the plasmid PUc8 solution with the resistant gene was added to the tube labeled positive. Gently give a few taps to the tube labeled positive to mix the plasmid solution. Both of the tubes were placed in an ice bath for 15 minutes. During the time period the tubes were incubating, two Luria plates and two Luria plates with ampicillin were retrieved. One of the Luria plates was labeled negative and the other positive, repeated on the luria plates with ampicillin. The group members’ names were then labeled on all of the plates. The cells of the bacteria needed to be heat shocked in order for the plasmid to enter into the cell. After removing both tubes from the ice bath, they were placed promptly into a water bath of 42 degrees celsius for 60-90 seconds. The tubes were then removed from the water bath and promptly placed back into the ice bath for two minutes. After this, the tubes were removed from the ice bath and 0.25 mL of the Luria Broth