How much of each bacteriophage dilution was added to

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μl. “How much E. coliwas added to each of the small Test Tubes after the Bacteriophage Dilution was added?” He says it was exactly 200 μl. “ Did he vortex each of the small Test Tubes after the Bacteriophage and the E. coliwere added and did he pour all 1.5 ml of Top Agar into the Bottom Agar Plate?” Yes. “ How did he create these serpentine Top Agars?” He says he poured the Contents of each Top Agar Tube into a Bottom Agar Plate. That’s it. He did not do a Mad Hatter Tea Party with any of these Plates. He simply poured the Contents of each Tube of Top Agar over the Surface of the appropriate Bottom Agar Plate and allowed the Top Agar to solidify in Place. “What are Alfonso’s Plaque Counts for each Plate of these Top Agar Slants?” 10-410-510-610-710-810-9TNTC TNTC TNTC TNTC 423 You look at Alfonsoʼs Plaque Counts, then you look at the Plates, and finally you tell Biff, Buckey, Buffy that their Phage Titer is probably -- 42
MIC 103L Exit Exam Form B 1 Page 1013. Some Virology Research Labs use Baby Hamster Kidney Cells (BHK Cells) to grow Mammalian Viruses in vitro. The Cells will grow as a Monolayer attached to the Collagen-Coated Bottom of a 75 cm2Flask (a 75 cm2 Flask usually contains approximately 15 ml of Growth Medium). To “splitʼa 75 cm2Flask (i.e. transfer a small Amount of Cells from the original 75 cm2Flask to several new 75 cm2Flasks) you need to determine the Total Number of Cells in the original 75 cm2Flask. • You remove all Media from the Flask and wash the Cell Monolayer attached to the Bottom of the Flask with 10 ml Phosphate Buffered Saline (PBS). You decant (“pour-off”) all of this PBS. • You add 5.0 ml of Trypsin-EDTA to the Monolayer and incubate the Flask for approximately 5 Minutes at 37°C. The Enzyme Trypsin and the Detergent EDTA will loosen and remove the BHK Cells from the Bottom of the Flask. You do not decant the Trypsin-EDTA. • You add 5.0 ml of Medium and transfer this Suspension of BHK Cells (now approximately 10 ml Total) to a 15 ml Conical Centrifuge Tube (just like the Green Cap Tube for the Hamburger Sample in Lab 6). • You vortex this Conical Centrifuge Tube and use a Microtiter Pipette to load a 5 μl Sample of this Cell Suspension into a Hemocytometer. Almost all Hemocytometers on this Planet are identical to the ones we use in MIC 101. Hemocytometers have two (2) Counting Chambers, each of which is divided (like “Hollywood Squares”) into Nine (9) Large Squares (1.0 mm x 1.0 mm). The Central Large Square is subdivided into 25 Medium Squares (0.2 mm x 0.2 mm). • You count the Cells in five (5) of the Medium Squares. - Your Counts are: 44, 39, 34, 46 and 37.

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