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Pablo Transformation Lab Report

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Mawjuda Abdulrashid Dr. Jesus Segovia Cell Biology Lab 3411L March 23, 2015 Transformation of E.coli with Plasmid DNA Abstract Transformation is the process by which DNA is introduced into bacteria. And then the bacteria can replicate and express that specific DNA. The purpose of the study was to study the theory of transformation and to develop an understanding of bacterial transformation by plasmid DNA. X-gal is a special sugar for the beta galacosidase enzyme and if we place this particular sugar into a bacteria, a very interesting thing will happen. “The ‘X’ in X-gal is a colorless pigment when attached to the galactose sugar but turns blue when cleaved by beta galaxtosidase” (Covert, 2011). Therefore, we observed blue colonies in our plate. There were approximately about 145 colonies present in plate A, 132 colonies in plate B, and no colonies in plate C. Our second data supports my hypothesis. There was a negative correlation between the fragment length and the distance migrated. The equation that we obtained was -0.0035x+51.319 and the r squared value was 0.8347. We grew E.coli bacteria in three different plates and observed whether they grew blue, white, or no colonies. This particular bacteria is not able to grow on ampicillin. My hypothesis is that cells exposed to LB (nutrient mixture) with ampicillin will …show more content…

Restriction enzyme basically are enzymes that cut DNA somewhere along its sugar phosphate backbone (Cheng 2015). They are naturally found in bacteria. Bacteria can be infected with viruses and as a way of protecting themselves against viral infections, what a lot of bacteria can do is that they can make these restriction enzymes. And the purpose of making them is to chop up any viral DNA that tries to infect the bacterial cell. The most important thing about restriction enzymes is that they cut the DNA at a particular and specific location, not random sequences (Nassan,

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Tn 4351 Unit 5 Lab Report

506 Words | 3 Pages

chinesis. A construct of R751::Tn4351 (the physical map of R751::Tn4351 and restriction sites are shown in fig. 7) was selected for introduction into F. chinesis to discover if the introduction and insertion of the vector R751 and the transposition of T4351 into the F. chinesis chromosome by a triparental mating occurred. One parent was E. coli GJ342 which carried a helper plasmid, the second parent was E. coli HB101 which contained R751::Tn4351 and the third parent was the F. chinesis target strain. 189 colonies were isolated on LB agar plates which in passage in fresh media were able to grow in 200µgml-1 erythromycin.

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How could a single nucleotide change on the plasmid alter the fragment pattern observed in the gel? (3 points) When a single nucleotide is change on the on the plasmid, it will make the restriction enzyme not to recognize the fragment pattern and also all the numbers at each site will be altered or this action could make the fragment not to be incised at the expected site, which could make a larger fragment to travels a short interval on the gel with less fragments than expected. It could also make it to be incised at more places than that originally expected and the result will be increased, smaller fragments that move further across the gel. There may also be no change in the banding pattern.

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Pglo Lab Report

1426 Words | 6 Pages

The other mediums were lacking all the necessary components for the activation of the GFP protein. The non-transformed E. Coli will be in the LB and LB + ARA plates since there is no ampicillin to kill

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Transformation Of Pglo Lab Report

984 Words | 4 Pages

Results Continued: The purpose of this experiment was to show the transformation of E. Coli with pGLO. We made four different plates each with different additives to compare them to one another, and be able to track the transformation. Initially we had predicted that only one of the four plates would glow. The plate with the plasmid (+pGLO)/LB/amp/ara was the one we said would grow and glow because it contained all the necessary tools to do so.

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Figure 3. Testing of transformed and mutant bacteria on minimal medium Growth was observed on the Transformed (Trsf) section and not on the Mutant (Mut)

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Pglo Transformation Lab Report

664 Words | 3 Pages

The hypothesis for this experiment was that transformed bacterial cells would grow on ampicillin plates and glow green when exposed to UV light. The rationale for this hypothesis was because the plasmid would code for ampicillin resistance and a green fluorescent protein, we would have the outcome explained in the hypothesis. 4. Our predictions were that for the standard protocol plates with agar and ampicillin, there would be growth and the colonies would glow under UV light. On the modified protocol on plates with agar and ampicillin where we changed the time of the heat shock, we expected less growth but the colonies would still glow.

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B-Galactosidase Lab Report

1764 Words | 8 Pages

B-galactosidase breaks down the disaccharide lactose into simple sugars glucose and galactose. However, glucose is a colorless compound hence it has to be substituted with a compound that is detectable by a visible color change. Hence,

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Pglo Transformation Lab Report

1057 Words | 5 Pages

In the presence of ampicillin, E. Coli growth will not occur. Therefore, in this experiment, bacterial growth on the plate indicates the uptake of the ampicillin resistant gene into the genetic code of E. Coli. If pGLO, LB broth, ampicillin resistance, and arabinose sugar are added, then the bacteria will glow. In one of the four plates that include all of those component, fluorescence will be observed. The purpose of this experiment is to demonstrate the genetic transformation that occurs in the E. Coli bacteria.

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Our hypothesis was that if the plate contains only the LB broth the E. coli bacteria would have no antibiotic resistance and would not glow. If the plate contains just LB broth and ampicillin then the E. coli bacteria will have antibiotic resistance only if the plasmid is present. If the plate contains LB broth, ampicillin and arabinose then the E. coli bacteria will glow fluorescent under a UV light and it will have antibiotic resistance. Similar to our expectations our results suggested that our hypothesis was correct. This is due to the fact that n order for the E coli.

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815 Words | 4 Pages

Introduction: This experiment includes the use of a genome editing system in order to make E.coli cells resistant to the antibiotic streptomycin. The system uses a protein called Cas9. Guide RNA is used to guide the Cas9 to the targeted sequence of DNA. The Cas9 finds the match to the gRNA in the cell’s DNA and cuts it out. As the cell tries to repair the DNA, it uses the template DNA that was inserted into the cell, and results in the modified DNA.

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806 Words | 4 Pages

In this experiment we investigated how a high or low mutation rate of E. Coli will affect its growth in a lactose medium and whether different concentrations of each of the strains would produce different results. We hypothesized that a higher mutation rate would result in more lactose digestion among the E. Coli cultures and that a higher concentration of either strain would produce more lactose digesting cells, and normal unmutated cells as well. As previously predicted, the results below show that the greater the plated concentration of E. Coli, the more mutated lactose digesting cells there were after a week of growth. However, one thing of note is that the low mutator plates especially the 10^3 concentration, produced more similar growth

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506 Words | 3 Pages

Hypothesis The transformed E.Coli with the ampicillin resistance gene will be able to grow in the ampicillin plates and it would have a green glowing color.

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coli strain W1485, which was one of the materials used in this experiment. Many techniques were used such as DNA isolation, transformation, and restriction enzyme digestion. Restriction enzyme digestion by EcoRI gave a linear fragment on the electrophoresis gel because the plasmid had only one restriction enzyme sequence site. The generated size of the plasmid was the same as the predicted size.

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The Pros And Cons Of Genetic Engineering

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The project has even served as a prototype for the Human Genome Project, illustrating the consequential impact such research could have on our world. As technology and knowledge of genetic engineering continues to advance, even larger and more ambitious projects can be undertaken; the research conducted on E. coli bacteria serves as the foundation and stepping stone for the small but substantial changes that can be made to any living organism - whether it be microbe, plant, or animal. The reengineering of such microorganisms has already been applied in testing for a cure to HIV, and this advance could soon lead to even more transcendent revelations. Consequently, the team is now seeking to recode the DNA segments of the synthesized E. coli into one unimpaired genome - testing whether the organism is capable of life and

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