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Pglo Lab Report

1426 Words6 Pages

Hypothesis: If the pGLO plasmid is inserted into E. Coli bacteria, then the surviving cells with the arabinose will glow green under UV light as they will be genotypically identical and phenotypically different. Experimental Design: In this experiment, recombinant DNA technology was used to modify the genetic properties of an organism. First a plasmid was created which consisted of ori, araC, GFP, bla and pBAD promoter. The GFP is a jellyfish gene that codes for the production of green protein; bla breaks down ampicillin which then can be used to select only the bacteria containing the bla gene by placing ampicillin in the growth medium. Ori is the origin of the DNA replication, araC makes a protein that binds to pBad promoter and switches …show more content…

The LB plate, the LB/AMP plate and the LB/AMP/ARA plate, all showed different phenotypic results under UV light. Only one of these plates glowed under UV light. The medium containing pGLO along with arabinose and araC was the only one that had a green fluorescence. The reason that this was the only one that glowed was because arabinose and araC activate the GFP protein. Only when arabinose binds to the AraC protein, the production of GFP is switched on. 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 …show more content…

The gel image, cells 9 and 10 show that they are both very similar in terms of genotype. They had almost the same band and similar content of RNA. The difference between the uncut pGLO band and our band was that the uncut went farther than our band. This could be because the uncut band is supercoiled and circular so it was able to move through the gel easier than the cut one. The cut band was linear and therefore ended up being slower. This also means that our band contains a restriction digest, where the uncut did not. This is because when DNA is supercoiled, it doesn’t have a restrictive digest and is able to move farther. The DNA HIND III digest showed 7 bands of different sizes. The pGlo Hind was smaller and therefore migrated faster than the linear EcoRI digest. The Uncut pGlo is supercoiled and therefore was able to migrate faster than pGlo EcoRI

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