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E. Coli Transformation Lab Report

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Gene expression using two types of plasmids (ampicillin resistance/lux), and their role in E. coli transformation. Nawaz Rahman Panther ID: 5029032 Signature:____________________________________ Lab Partners: Manuel Vera Giselle Janoura Jeniffer Marranca Section U17 Abstract Small circular pieces of DNA molecules located inside the nucleoid in bacterial species (prokaryotes) are known as Plasmids.Plasmids do not dictate the survival of the host bacteria, but it can give rise to certain genes being activated, if required, providing certain benefits. Survival and reproduction in different environments can be considered as one of these benefits. In this experiment we add different …show more content…

Once the membrane has been chipped away by the calcium chloride it is extremely important to heat shock the bacteria, which is essentially to create large gaps in the membrane so the plasmids can enter the E. coli (Goldina et.al 2011). E. Coli will be grown in round plates already containing agar. Agar is a gel like substance that the E. Coli will use to feed on and grow. Luria broth is another important part of the experiment, it provides nutrients E. Coli needs in order to grow. Luria broth can be considered as cow manure. Luria broth is so effective in keeping the E. Coli alive that it is able to adapt to different aerobic or anaerobic incubation conditions in order to sustain its life (Semenov et.al 2011). This experiment can be used as a basic model for the basic principle, which is when information in the DNA is converted to an RNA copy by means of transcription then having the RNA copy directing the assembly of a chain of amino acids through translation (Raven 2011). Scientists can use biotechnology to manipulate plant genes resulting in the invention of pesticides. …show more content…

After making them competent the cells are heat shocked to open up the membrane pores for the plasmids to slide in. Next segregation takes place where the E. Coli cells are placed into different tubes where the rest of the substances are added. Three out of six of the petri dishes resulted in lawn growth, meaning E. Coli was able to grow freely. The petri dish containing E. Coli, Luria Broth and ampicillin resistance gene resulted in lawn growth. Nothing in this petri dish could hamper the growth of E. Coli, hence it was destined for lawn growth. The second dish to display lawn growth contained the same substances as the one mentioned above, except it did not have any plasmids. The results were the same because no ampicillin was inserted in here, just like the previous one, allowing E. Coli to replicate freely. The final petri dish to have lawn growth contained Luria Broth, E. Coli and the lux gene. The lux gene should allow the bacteria to glow but it failed to do so. This is because the newer E. Coli cells do not contain the lux plasmid and are taking over the older E. Coli cells. Two out of six of the petri dishes displayed colonial growth. The first one contained E. Coli, ampicillin,

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