Shake Flask Fermentation
In this experiment, Escherichia coli is used as a sample to study the growth kinetic of microorganism in shake flask. A different volume of E.coli was transferred into 250ml Erlenmeyer/shake flask containing media for the nutrient of microorganism. The different volume of microorganism transferred will give the different effect of reading on the constant volume of media used. There are three ways to test the growth kinetic rate of microorganism on shake flask, which are by optical …show more content…
Cell lysis occurs in this phase, and growth can be re-established by transferring to fresh media.
5.0 Apparatus * Microbe on agar plate (E.coli) * Shake flasks * Sterile loop of wire * Cotton plug (cotton, cotton mesh,aluminium foil) * Incubator shaker * Media for inoculums growth (LB) * Pippettor * Centrifuge tubes * Cuvette * Falcon tubes * Light (bunsen burner) * Centrifuge * Ethanol * Distilled water * Sterilized graduated cylinder
6.1 Media preparation 1) Luria Bertani (LB) broth (Lennox) is calculated to get concentration for 200ml of broth. LB brpth composition written to make media on the box that stored the LB is 10 g/L. So, 1.6g of LB broth powder is diluted with 160ml of distilled water to make the media. 2) 160ml of the LB broth is prepared inside 250ml Erlenmeyer flask. 3) Erlenmeyer flask is closed with cotton covered by gauge and aluminum foil. 4) 10 g/L of glucose is diluted with the same amount of distilled water used to dilute media that is 160 ml and is placed in a schott bottle. 5) The Erlenmeyer flask containing media with all the experiment is autoclaved for 3 hours.
6.2 Sampling for absorbance analysis/optical density
1) 2 ml of inoculums is taken out and being transferred into cuvette. 2) 2 ml of blank (LB broth not contain microorganism) is transferred into cuvette. 3) The spectrophotometer is