The purpose of this lab is to use genetic engineering to transform E. coli bacteria by inserting

This pBlu lab had for purpose to present the changes of the strain of E. coli bacteria due to new genetic information being introduced into the cell. In this experiment we are freezing and heat shocking the E. Coli bacteria that is then forced to take the plasmid DNA. The E. coli then transforms the pBLu plasmid, which carries the genes coding for two identifiable phenotypes. After following the Carolina Biological steps our lab worked well and we able to see some colonies of bacteria on the plates. The x-gal plate showed a significant amount of bacteria to confirm that the pBlu plasmid took over the E. coli strain.

E. coli HB101 was transformed with pGLO plasmid then grown on media containing ampicillin and/or arabinose and on medium containing neither (Brown, 2011). This is done for selection of transformed cells since not all cells are expected to take up the plasmid (Brown, 2011). We also expect roughly the same CFU on any plate(s) receiving samples from the same microcentrifuge tube, since they are getting the exact same

This experiment was designed to test and observe the transformation efficacy of the pUC18 and lux plasmids in making E. coli resistant to ampicillin. Both plasmids code for ampicillin resistance, however, the lux plasmid codes for a bioluminescence gene that is expressed if properly introduced into the bacteria’s genome. The E. coli cultures were mixed with a calcium chloride solution and then heat shocked, allowing the plasmids to enter the bacteria and assimilate into the bacterial DNA. The plasmids and the bacteria were then mixed in different test tubes and then evenly spread onto petri dishes using a bacterial spreader, heating the spreader between each sample to make sure there is no cross contamination. Each of the dishes was labeled and then incubated for a period of 24 hours. The results were rather odd because every single one of the samples grew. Several errors could have occurred here, cross contamination or possibly an error in preparation as every single sample in the class grew, meaning all samples of the bacteria transformed and became ampicillin resistant.

In the pGLO Bacterial Transformation lab, Escherichia coli is transformed with a gene encoding green fluorescent protein by inserting a plasmid containing the GFP gene, beta-lactamase, and arabinose into the bacterium. Successfully transformed bacteria will grow in the presence of ampicillin and glow a bright green color under ultraviolet light. The sugar arabinose is responsible for switching on the GFP gene in the transformed cells, without it, the gene will not be expressed.

The petri plate with the +pGLO LB/amp/ara will allow for the E. coli who have accepted the plasmid to glow and express the GFP. The E. coli will only express GFP if they are exposed to the arabinose sugar, and the bacteria also have to be exposed to the pGLO. Also the bacteria that are exposed to the ampicillin, but are not exposed to the pGLO will perish. Other plates may grow, but not express the protein.

After analyzing the data recorded for both the agar plates containing ampicillin and those that did not, it can be concluded that the data provides enough evidence to reject the null hypothesis. There is enough evidence to support the alternative hypothesis stating that there is a correlation between plasmids coding for an antibiotic resistant gene and bacterial growth in ampicillin. When a bacterial solution containing either pUC18 or the lux plasmid is transformed in an agar plate containing ampicillin, only those cells which took in the plasmid are able to survive and replicate, forming individual colonies. Not all cells are transformed though, the chances of a successful transformation were extremely low. In order to see which cell transformed the cells were tagged according to their plasmid, in the plates containing pUC18 the only

In preparing for the bacterial transformation, DNA plasmid is introduced into the E. coli cells that will express newly acquired genes. Two tubes were used and labeled both as +pGLO and -pGLO. A solution of (CaCl2) was transferred 250 µl onto the two tubes. The tubes were placed on the ice. A sterile loop was then used to gather a single colony of bacteria from a starter plate. Now, that both tubes contain bacteria they were placed on the ice for 10 minutes. Four agar plates were labeled as: +pGLO LB/amp, +pGLO LB/amp/ara, +pGLO LB, -PGLO LB/amp. Heat shock was used to transfer both the +pGLO and -pGLO, at exactly 42°C. Time was observed for 50 seconds and quickly return the tubes to the ice for another 2 minutes. As the tubes, cold down they

a. LB+plasmid and LB-plasmid: Both of these plates had a lawn of bacteria. This proves

While leading this analysis, gene expression was critical to the results of the non-transformed and changed cells. In its procedure of choice, the bacterial cells should have picked if they ought to allow the ampicillin safe quality, therefore indicating commendable components of quality expression. The Escherichia coli's cells were not changed, there seems to have been an error during the experiment. There should have been a slight change in a way that there should have been a colonial growth design in the bacterial cells that had ampicillin safe plasmid. (Hidalgo, Nuria Fernández) On the other hand the cells that were on the ampicillin (lux, control, and NP) demonstrated a colonial design because of the non-transformed cells dying due to

This experiment was completed to conduct bacterial transformation and introduce ampicillin resistant DNA plasmid to E. coli bacterial cells. Observations are made of the difference in cell growth due to various environments provided within agar

According to our lab manual, Genetic transformation is when the genetic makeup of an organism is altered by incorporating external genetic material (Barnhart and Hopper 2014). Genetic transformation was discovered on accident by the scientist Frederic Griffith in 1928. He was trying to create a vaccine for pneumonia, but instead discovered bacterial transformation (Barnhart and Hopper 2014). Plasmids, which are a genetic structure in a cell that can replicate independently of the chromosomes, are used as a form of extra genetic material in order to transform bacteria. They are expressed by both the bacterial cell and the daughter cell. In the case for our genetic transformation lab, our cells we are using are E. coli cells, and

This experiment focuses on genetic engineering and transformation of bacteria. The characteristics of bacteria are altered from an external source to allow them to express a new trait, in this case antibiotic resistance. In is experiment foreign DNA is inserted into Escherichia coli in order to alter its phenotype. The goal of the experiment is to transform E. coli with pGLO plasmid, which carries a gene for ampicillin resistance, and determine the transformation efficiency. The bacteria are transformed by a combination of calcium chloride and heat shock. When the bacteria are incubated on ice, the fluid cell membrane is slowed and then the heat shock

The purpose of this lab is to observe bacterial growth under various conditions including the transformation of bacteria; to understand how the process of transformation occurs.

Ampicillin is a beta-lactam antibiotic which can be used to treat a large number of infections. For example, Escherichia coli (E. coli) bacteria is terminated by this specific antibiotic. Ampicillin interferes with the formation of bacterial cell walls and thus kills newly dividing cells that must form new cell walls. Plasmids contain genes that create antibiotic resistance to their host cell. The pGlo plasmid contains an Ampicillin resistance gene. Therefore, bacteria that take up the plasmid and transform become resistant to Ampicillin. To carry out this experiment, my colleagues and I took four petri dishes containing the bacterial host cells (E. coli) on each and the Ampicillin on two of those four. Through a tedious process, we added the pGlo plasmid to one petri dish of just the host cell and one petri dish of the host cell and the Ampicillin antibiotic. When examining the results of our experiment, we noticed that both petri dishes containing zero Ampicillin entirely submerged in a lawn of the Escherichia coli bacteria, the petri dish containing no plasmid but some Ampicillin displayed zero growth, and the petri dish containing the plasmid and Ampicillin showed individual colonies of the E. coli bacteria glowing under a UV light. The reason this occurred is because the petri dishes that showed a lawn of bacteria contained no Ampicillin so there was no antibiotic to kill the bacteria off. The petri

This experiment was performed to test the hypothesis if LB nutrient broth, +pGLO and -pGLO Ampicillin, and Arabinose was placed in the E. coli plates, then there will be a significant growth in the newly transformed bacteria and it will possess the ability to glow under UV light. The measurements were recorded from the bent glass tube in each glass test tube. The transformation protocol tested for the newly possessed traits in E.coli bacteria. Throughout the experiment there were many probable reasons for failure. If the pipettes and sterile loop were not thrown out in between each use, a cross contamination could cause a miscalculation in the experiment causing the data results to fail. The hypothesis that was tested was validated due to the positive results with each experiment stating that newly transformed organisms due in fact pass on traits.