Pglo Report

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Quality Report on pGLO prep
Genetic transformation is a process that modifies bacteria, by introducing new genetic material. In our lab we introduced the pre-engineered pGLO into the E. Coli HB101 K-12 bacteria. This pGLO plasmid consists of the gene for the green fluorescent protein (GFP), the ampicillin resistance gene that inactivates the ampicillin in the LB media and the araC gene that indirectly controls the arabinose digestion enzymes [Fig 1].

Green Fluorescent Protein fluoresces bright green when exposed to UV light. The GFP gene only activates if there is arabinose present. When arabinose is not present, the arabinose digestion genes are inactive and energy will be conserved. However, when arabinose is present the genes activate and start to break down the arabinose until it is all consumed. BAD encodes for the enzymes used to digest arabinose. The araC gene in the DNA map above pairs with arabinose, and up regulates BAD. However, it also negatively feeds back into its own regulation. Bla is the Ampicillin Resistance gene produces Beta lactamase, a protein that confers ampicillin resistance [1].

In our lab, we used chemically competent cells, which are bacteria cells that …show more content…

A ratio of approximately 1.8 is considered pure for DNA. If the ratio is lower than 1.8, it indicates some contamination. This contamination can be attributed to the presence of protein. Another measure of purity is the ratio of absorbance at 260 and 230nm. This ratio is generally slightly higher than the 260/280 ratio and a ratio of approximately 2.0-2.2 is considered to be pure. In this case, if the ratio is lower than 2.0, it indicates contamination [3]. Our Nano-drop data after we purified the plasmid came out to be 1.91 and 1.42 respectively. Our 260/230 ratio was significantly lower than our 260/280 ratio that signifies that there was some other contamination in our

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