03_Lab 2 - Lab2.Gramstain&Streakingforisolation...

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Before we start…. -  Lab manual and the kit should be on the table. -  Put your backpack on the metal shelf (NO FOOD in lab) - Tie your hair -  Turn in the lab 1 report (before 6:10 pm) -  Wipe the bench with disinfectant spray/wash your hands
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Safety  Refill  the bottles if necessary – be careful with Gram 1: will not  come out of clothing, hands etc. Use taps from benches if you need to refill the water bottle     Be careful when you turn on the taps! (very high pressure!) Do not use hand-washing sink for gram staining!  2. Gram stains 1. Burner Two students share one burner. Do not leave the burners without attention –  whenever you get  up from your seats, you should turn off the gas jet.  If the flame is found unattended, 5 points will be off from  technique/participation.  Keep flame away from gram stain bottles –Gram III is ethanol! Turn it off whenever it is not in use. If your flint is not working, please ask TAs to change it. Before you leave, check if the gas get is turned off. 
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5. Dispose correctly 3. Test tubes Always carry culture tubes in test tube racks (2~4 people/rack).  Never take out the rack (white color) from the incubator. 4. Microscope (in the cabinet) No glass slides in the orange and red biohazard bags! Contaminated?   Orange or red biohazard bags. Orange bag should be empty out before you leave        dump the content into the red bag.  Stage holder should not be fully extended. 4X object should be on the stage.  Show TAs your microscope before put it back. Safety 
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Things to know for Lab 2 2. Pure culture  A culture that contains cells of one kind, all progeny of a single cell  1. Aseptic technique (sterile technique) The process by which one handles materials without introducing  unwanted microorganisms. cf)  Colony forming unit (CFU) vs. colony      -  CFU is a single living bacterium that can grow to form a  colony      -  Colony is a cluster (or progeny) of a single cells
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Things to know for Lab 2 3. Culture media Chemically defined (synthetic) medium  Composed of ultra-pure organic  /inorganic compounds  Time-consuming to prepare  Useful to study the minimum  nutritional requirements of a  particular microorganism or  metabolic activities.   Little value in diagnostic  bacteriology Complex medium  Supply an excess of all  nutrients  Easy to prepare  Hard to know the exact  chemical composition  Permit many organisms grow
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Example of complex media Component Amount
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This note was uploaded on 02/18/2010 for the course PHYS 317L taught by Professor Lang during the Spring '10 term at University of Texas at Austin.

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03_Lab 2 - Lab2.Gramstain&Streakingforisolation...

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