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Aseptic Germ Theory - Biology 151 1 LABORATORY MODULE 3...

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Biology 151LABORATORY MODULE 31Aseptic Technique & Germ Theory of DiseaseAll living cells share many biochemical and physical similarities; however not allcells are equivalent.Prokaryoticcells do not contain a membrane-bound nucleus like theeukaryotes you have seen.Instead their DNA is suspended in a portion of the cytoplasmcalled thenucleoid.Prokaryotes are also typically much smaller than eukaryotes.Eukaryotic and prokaryotic cells exhibit other important structural and chemicaldifferences.Many of these differences determine the role of prokaryotes in nature, fromrecycling nutrients to causing disease in humans.The differences between eukaryotic andprokaryotic cells also provide the basis for treating diseases caused by bacteriaGrowing MicroorganismsTo culture microorganisms, or any cell type, the cells must be provided with amedium containing essential nutrients and environmental conditions that are consistentwith maintaining life.Bacteria can be maintained as a culture in either a liquid media(broth) or on a solid media (agar plates). Agar is a gelatinous material derived from certainmarine algae used as a base for bacterial culture media. Some media such asTSA(Trypticase Soy Agar) orTSB(Trypticase Soy Broth) contain a rich supply of nutrients andwill support the growth of most types of bacteria.Aseptic Techniques – Some General Rules1. Disinfect the bench top with an appropriate disinfectant before you begin working andafter you are finished with the experiment.2. Work on a clear bench top.Put away all unnecessary items.3. Wear the necessary personal protective equipment (lab coat, gloves, goggles, etc)4. Wash your hands before and after manipulating a sample.5. Keep all cultures closed and tubes upright in a rack until ready for use.6. Hold cultures by the tube NOT the cap.7. Work quickly and carefully - don't become distracted or allow others to disturb you.8.Waste Disposala. Contaminated paper & plastics go in the biohazard trash bin.b. Contaminated glass (with or without bacteria) NEVER goes into the biohazardtrash bin – it goes into the contaminated broken glass bin in the fume hood.c. When in doubt – ASK!
2LABORATORY MODULE 3What is Aseptic Technique?Most experiments performed using microorganisms usesterilemedia andpurecultures(progeny of a single organism).A series of operations have been developed tolimit the risk of contaminating these materials during manipulations.These operations areknown asaseptic techniques.They also help protect investigators from infectingthemselves or others by preventing the release of the organisms into the environment.When working with any cell culture it is critical that you know the rules and proceduresand understand how each requirement contributes to maintainingasepsis(state of beingfree of living pathogenic microorganisms).

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Term
Fall
Professor
Sarah Daoudi
Tags
Biology, molecular biology, loop, Agar plate, Petri dish, sterile inoculating loop, Microbiological culture

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