Clostridium Perfringens Lab Report

Introduction
The three organisms that purification was performed on were Clostridium perfringens, Marinithermus hydrothermalis, and Desulfovibrio hydrothermalis. M. hydrothermalis comes from a deep-sea hydrothermal vent, it has rod shaped cells and is gram-negative. It grows at an optimum temperature of 67.5 C, an optimum pH of 7, and a salt concentration of 30% (1). D. hydrothermalis comes from a deep-sea hydrothermal chimney. It is vibrio shaped and gram-negative. D. hydrothermalis has an optimum temperature of 35 C, an optimum pH of 7.8, and in a salt concentration of 2.5% (2). C. perfringens is a rod-shaped, spore forming, gram positive bacteria. It grows at an optimum temperature of 37 C and optimum pH of 7-7.5 (3). It is found as part …show more content…
The M. hydrothermalis plasmid was prepared from overnight culture, digested by restriction enzymes, expressed with pET system expression, purified through a nickel histag affinity column, and ran on an SDS-PAGE gel. The C. perfringens DNA was transformed, its plasmid was prepared from overnight culture, purified using affinity chromatography, and ran on an agarose gel. The D. hydrothermalis plasmid was prepared from overnight culture, purified using affinity chromatography, and ran on an agarose gel. Each sample was quantitatively measured using Bradford and kinetics assays. Characterization was performed on D. hydrothermalis because it was the only organism that had activity in the kinetics assay after …show more content…
Agarose gels are constructed out of solid agarose and 1X TAE buffer. After the agarose has been melted in the buffer, 0.5 ug/mL ethidium bromide is added to the liquid. This liquid is poured into a gel box and left to solidify. 0.5 ug/mL ethidium bromide is placed on top of the solidified gel and 1X TAE buffer is poured over the top to suspend the gel in buffer. 2 uL of Bromophenol blue/xylene cyanol tracking dye is added to each sample and these are loaded into the wells of the gel. The gel runs at 100V for about an hour. After the gel has finished running, it can be examined over a

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