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All slides go into the stainless steel pitcher you ʼ

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• All Slides go into the Stainless Steel Pitcher You ʼ ll probably be giving a Buttload of Plates the Heave-Ho at the End of Today ʼ s Lab. 谢谢 ! Xie-Xie! _______________________________ “So it's true? You've found Nurhachi?” ‘You know I did. Last night one of your boys tried to get Nurhachi without paying for him’. “You have insulted my son.” ‘No, you have insulted me. I spared his life.’ © 1984 LucasFilm, Ltd.
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Lab 6 Page 4 Lab 6 Exercises Viable Count Materials (per Clan): • Six 16mm Sterile Test Tubes • Test Tube Rack • 1 Bottle of Sterile Saline • 5 ml Serological Pipettes • 6 Nutrient Agar Plates • 1 Packet of Spreaders • 6 MacConkey Agar Plates • 1 Turntable • P1000 Microtiter Pipette (and Tips) • Bunsen Burner • P200 Microtiter Pipette (and Tips) The Basic Set-Up: 1. Label all your Test Tubes by writing on Tape with your Sharpie and then attaching this (temporarily) to the Test Tubes. • Your Test Tubes really ought to be labeled: 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 and 10 -7 • But you can get away with labeling them: 2, 3, 4, 5, 6, and 7 2. Label your Nutrient Agar Plates by writing directly on the Plates • Your Nutrient Agar Plates really ought to be labeled: 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , and 10 -8 • But as with the Phage, it’s less Ambiguous to label them: 3, 4, 5, 6, 7, and 8 Plates are labeled one Dilution Factor off from the Dilution Tubes. We ʼ re only transferring 1/10 ml to each Plate so we need to “up” the Dilution Factor by a Factor of 10 on Labels for the Plates (so a 10 -7 Dilution Tube Sample goes onto a 10 -8 Plate). It ʼ s just like with the Phage Titer Experiment. 3. Label your MacConkey Agar Plates by writing directly on the Plates • Your MacConkey Agar Plates should be labeled: 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , and 10 -8 • But again, let’s just label them: 3, 4, 5, 6, 7, and 8
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Lab 6 Page 5 Set-Up for Dilution Series (Sterile Technique) 1. Using Sterile Technique, add 4.5 ml of Sterile Saline to each of 6 Large Test Tubes that you have put in your Test Tube Rack. A. Loosen the Lid on the Bottle of Sterile Saline Don ʼ t remove the Cap; just loosen it. B. Open your sterile 5 ml Serological Pipette at the End of the Package (where the Packet states, “Peel Here To Open”). Open the Wrapper but don ʼ t remove it yet. C. Attach a Lightsaber to this sterile Serological Pipette . Seating the Pipette in the Rubber Cone inside the Lightsaber takes quite a bit of Force. D. Once the sterile Serological Pipette is seated in the Lightsaber, raise the Plunger on the Lightsaber about a Centimeter. This gives you enough Volume to “blow-out” all of the Tryptone Broth (remember, these are TD [To Delivery] Pipettes). E. Open the Wrapper a bit more and remove the Pipette. Keep the Wrapper! F. Remove the Lid from the Sterile Saline Bottle and use the Wheel on the Side of the Lightsaber to aspirate (“suck-up”) 4.5 ml of Sterile Saline.
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