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2 use a separate sterile toothpick to transfer a

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2. Use a separate Sterile Toothpick to transfer a Goober of our Negative Control (Streptococcus lactis ) onto the Right Side of the same Slide. 3. Add a Couple of Drops of Hydrogen Peroxide (Clear) to each Smear. 4. Either Skin Isolate Species should start fizzing like Wombat Spit. The Reaction will usually occur within about 15 Seconds at the most. 5. Streptococcus lactis will do nothing. It will look as though you added a Couple of Drops of Water. Please put used Toothpicks in the Microtiter Pipette Waste Beaker on your Bench. Please transfer your Toothpicks and Slides to the Stainless Steel Pitcher on the Waste Cart. Yeast Glucose Calcium Carbonate (YGC) Plates Materials (per Clan) • 1 YGC Plate • Your Skin Isolate (Micrococcus luteus or Staphylococcus epidermidis ). Procedure 1. Streak your Skin Isolate onto a new YGC Plate. 2. Incubate at 37°C for 48 Hours
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Lab 6 Page 13 Gram Negative Biochemical Tests Oxidase Materials (per Clan): • Sterile Toothpicks • Filter Paper Disks • Oxidase Reagent (Light Blue Stuff in the Ice Bath on the Back Bench) • Our E. coli and our Pseudomonas putida • Your Pseudomonas sp. Isolates (Each Member of the Clan should have their own individual Pseudomonas Isolate) Oxidase only makes Sense when it is performed on Gram Negative Bacteria. Procedure 1. Use a Sterile Toothpick to transfer a Goober of our Positive Control (Pseudomonas putida) to the Left Side of a Disk of Filter Paper. 2. Use another Sterile Toothpick to transfer a Goober of our Negative Control (E. coli ) to the Right Side of this Disk of Filter Paper. 3. Add a Couple of Drops of Oxidase Reagent (Light Blue) 4. The Goober of Pseudomonas putida should turn dark Blue within about 10 Seconds. The Goober of E. coli should remain that vaguely nasty Latex Color for at least 30 Seconds (but be aware that both Goobers will turn Dark Blue after a few Minutes.) 5. Repeat the Above, using a Sterile Toothpick to transfer a Goober of our Pseudomonas putida to the Left Side of a Disk of Filter Paper, and another Sterile Toothpick to transfer a Goober of your Pseudomonas to the Right Side of this Disk of Filter Paper. The Oxidase Test is the most frequently misinterpreted Biochemical Test. Errors in interpreting the Oxidase Test can make you falsely assume you have an Enteric or a Pseudomonas . This can cost you 20 Points on the Biochemical Unknowns. Please put used Filter Paper in Oscar (not in the Trash). Please put used Toothpicks in your Microtiter Pipette Waste Beaker, and please transfer them to the Stainless Steel Pitcher on the Waste Cart.
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Lab 6 Page 14 F Agar Materials (per Clan): • 4 F Agar Plates (Brown Stripe [1 per Student]) • Inoculating Loop • Bunsen Burner • Your Pseudomonas sp. Isolate (Each Member of the Clan should have their own individual Pseudomonas Isolate) F Agar should only be used to confirm that a Gram Negative, Oxidase Positive Bacterium is Pseudomonas .
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