Thursday, March 6, 2014

PGLO TRANSFORMATION LAB

Purpose:The purpose of the lab was to show the process of transformation. With that concepts we were testing was the the effects of ampicillian and the sugar arabinose had on the growth of the cells. The relationship between the dependent variables was that we measuring the growth of cells would differ depending which plates had the ampicillian which the pglo was resistant to. For the independent variable we had our control group as the plate with just broth and no ampicillian and the others will be the ones with the varied results. What were trying to find out the success of transforming the gene into the organism E-coli given the variables.

Introduction: The lab will be conducting a process which is known as genetic transformation caused by genes. Gene transformation will is basically inserting a trait from an organism to another. The trait that will be expressed is GFP (Green Fluorescent Protein). If the transformation is successful then the organism which is E-coli in this case will glow under ultraviolet light. For the gene to go off, the sugar arabinose is added to the cell. Cells that have transformed will grow on the plates that have lb/amp because the pglo is resistant to the antibiotic ampicillin.


Methods: During the lab, our group would carefully read out the measurements just in case to confirm, what we were doing was correct with the other lab partners.. We made sure that we recognized each plate and continued accordingly. In between tasks if we had the opportunity, we would answer the questions pertaining to the lab. It was easy to answer them because it was either still fresh in our minds or predictions about the lab. Of course we couldn't answer the end questions because we had to wait 24 hours to answer those. At the end to make sure that we were successful we went over the plates until it was covered in transformation. Overall the lab procedure was clear and simple and we adjusted to what was more convenient for us and that was to organize and break up the lab equally to be more efficient.

Data:


Graphs and Charts: (see above)

Discussion: The normal e. coli culture with no pGLO introduced had high e. coli growth on it, as would be expected; additionally, the normal culture that had been inteoduced to ampicillin had no growth due to ampicillin's role in stopping bacterial growth. These two are the control plates, so it's to be expected that they behaved the way they normally would. The transformation plates contained the most important data. With the pGLO introduced, its natural resistance to ampicillin was present, so those colonies of e. coli which were transformed by the pGLO managed to survive even with ampicillin present, resulting in a few surviving colonies. In the final culture, arabinose was added to the culture, and because arabinose stimulates the GFP to cause phosphorescence, the colonies found in that culture did glow under ultraviolet light.

Conclusion: There wasn't really a definite answer to the lab however, it was a success in having E-coli taking on the gene that made it glow green under fluorescent light. What went wrong during the lab was the outcome or in the lab stated as the efficiency of the about of cells that actually took on the trait. We had a very low outcome in the end but was successful.

References: The lab itself


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