Lab Report Essay

Lab Report Essay
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Transformation of E. coli with Green and Blue Fluorescent Protein Plasmids.
Introduction
Bacterial transformation involves the ability of the bacteria to obtain as well as portray new traits by fusing foreign DNA into their cells through their cell walls (Science Bridge 6). According to Ontario Science Centre, bioluminescence is a biochemical process that enables living things such as fungi, insects or marine fish to produce light. For instance, Jelly fish Aquorea Victoria emits green lighting thanks to the green photogenic protein located in unique photogenic cells at the base of its umbrella (13-14).
As observed by EDVOTEK (5-6) this exciting ability of bioluminescent marine organisms has inspired scientists to genetically engineer blue fluorescent protein (BFP). The protein contained in green fluorescent protein (GFP) is combined with other proteins to study different biochemical processes in cell and molecular biology experiments. GFP and its variant BFP can be used together to show the impact of pivotal amino acid transformations with regard to the structure and functionality of a protein. This will help learners understand illnesses that are genetically transmitted, as well as the impacts of several mutations on aging and when different diseases begin. The objective of the experiment was to gain knowledge of the biological process that involves the transformation of bacterial cells by pFluoroGreen or pFluoroBlue DNA. From the experiment, I was able to observe the acquired phenotypic trait of green or blue fluorescent protein exhibited by transformed bacterial cells.
Material and equipment used
With advice from EDVOTEK (3-4), I used the following materials and equipments during the experiment: Transformation cell slant, CaCl2, ReadyPour medium and Luria Recovery Broth which were stored at room temperature. The following were stored under freezer conditions; super coiled pFluoroGreen, supercoiled pFluoroBlue, ampicillin and IPTG. Other equipments included two petri plates (small and large), plastic microtipped transfer pipets, sterile wrapped 10 ml pipet and sterile wrapped 1 ml pipet. I also used sterile toothpicks, sterile inoculating loops and microtest tubes that had attached lids. Several requirements for the experiment include: Automatic Micropipet (5-50 μl) and tips, two water baths stored at 37°C and 42°C respectively, thermometer, two incubation ovens at 34°C and 42°C respectively, pipet pumps, ice, marking pens, Bunsen burner, hot gloves and long wave U.V. light.
Procedures basing on EDVOTEK (13 -16)
Setting up the transformation and control experiment.
Mark one microcentrifuge tube “+ DNA” which is the transformation tube, and another one mark as “– DNA ” which will be experimental control tube.
Add 0.25ml of ice cold CaCl2 to each tube using a sterile 1 ml pipet.
Transfer colonies from the source plate of E. coli cells. For the test tubes labeled “+DNA” and “–DNA”, apply these methods: Use a sterile toothpick to transfer 2-4 colonies from the source plate to the test tube. Between your fingers, twist the tootpick vigorously and up and down in the cold CaCl2 solution to dislodge the cells.
Suspend the cells completely in both tubes by tapping.
To the “+ DNA” tube, add one of these: 10 μl of FluoroGre...