Bugbuster Lysis Synthesis Lab Report

The experiment consists of 5 steps—Inoculation, fermentation, cell lysis, inclusion body solubilization, and peptide purification. In the first two steps, E coli with Snake 6 peptides is cultivated. Ultraviolet–visible spectroscopy (UV-vis) is used to quantify bacteria through detection of their optical density (O.D.). After reaching a certain concentration in the fermentation batch, inclusion bodies (IBs) are extracted from these E coli by adding Bugbuster lysis solution and lysozyme. Polyacrylamide gel electrophoresis (SDS-PAGE) is carried out to test the existence of IBs. Afterward, IBs are dissolved in 8M urea and poured into IMAC chromatography in peptide purification step. Optimization of each step is tested throughout the experiment. Table 7 below shows the experimental matrix for the experiment.
with Snake 6 would be obtained from Dr. Claire Komives. Media in the fermentation shown in Table 2 and all chemical solutions would be obtained from San Jose State University (SJSU). Glucose meter would be used to detect glucose concentration. The incubator would be used for inoculation. Bugbuster lysis solution, lysozyme, urea, and other chemicals would also retrieved from SJSU. Centrifugation would be carried out through RC-5B Refrigerated Superspeed Centrifuge. In chromatography, IMAC column would be used for peptide purification. UV-vis and SDS-PAGE would be our analytical tools. All equipment would be provided by