Lab5-2011

Lab5-2011 - BIOL 513 – Lab 5 Virus characterization –...

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Unformatted text preview: BIOL 513 – Lab 5: Virus characterization – Part 3. Thermostability and pH sensitivity 78 Lab 5: Virus characterization – Part 3. Thermostability and pH sensitivity- lab designed for 12 groups Equipment for Lab 5: Day 1 Day 2 Day 3 Day 4 Day 5 • CO 2 incubator + + • 6-ft biosafety cabinet - 2 • Light microscope - + • Inverted phase contrast microscope + - • Hemocytometer with coverslip - 12 • Hand counter - 12 • Vortex 6 - • 2-liter vacuum flask with aspirator 2 + • Pipetman, 2-20- µ L 12 - • Pipetman, 40-200- µ L 12 12 • 8-channel Pipetman, 50-300 µ L - 6 • Safety bulb 12 12 • 250-mL beaker 12 - • Forceps - 12 • Rack for microcentrifuge tubes 12 12 • Rack for 12x75-mm tubes 24 - • Rack for 50-mL tubes 12 12 • 56°C water bath & rack + - Supplies & Consumables for Lab 5: Day 1 Day 2 Day 3 Day 4 Day 5 • 96-well tissue culture plate cluster - 24 • Microcentrufuge tube - 12 • Sterile 12x75-mm tube with snap cap 576 - • Sterile dilution trays - 24 • 1-mL pipet 72 - • 10-mL pipet - 26 • Sterile Pasteur pipet 24 6 • Sterile filtered pipet tips in trays 12 12 • Yellow pipet tips in trays - 12 • 0.4% Trypan blue stain - + • Crystal violet stain in 50% methanol - + • Glycine-HCl buffer, pH 3.0 + - • 70% ethanol in spray bottle 6 6 • Paper towels - + • KimWipes - + Also check the corresponding sections of Lab 6, which begins on day 2 of Lab 5. BIOL 513 – Lab 5: Virus characterization – Part 3. Thermostability and pH sensitivity 79 Lab 5: Virus characterization – Part 3. Thermostability and pH sensitivity- lab designed for 12 groups Materials for Lab 5 provided by the instructor: Day 1, Tuesday, March 1, 2011 • Samples of FCV and FHV, 0.5 mL at 5x10 6 PFU/mL each – 12 sets • 2 M NaOH, 100 µ L in 1.5-mL tube – 12 • Glycine-HCl buffer, pH 3.0, 2 mL in 12x75-mm tube – 12 • 50-mL tube with 50 mL CrFk cell culture medium (DMEM +10% FCS, 1x AA) – 12 • Extra 96-well plate cluster with seeded cells – 6 • Extra CrFk medium Day 2, Monday, March 7, 2011 • 24 mL suspension of freshly trypsinized CrFk cells @ 4x10 5 cell/mL – 12 (from 5-6 150- mm plates @ ~2x10 7 cells/plate to get total 120x10 6 cells) • Extra CrFk medium Also check the corresponding sections of Lab 6, which begins on day 2 of Lab 5. [Instructors: for every 2 lab groups, you will need to prepare 1 12-well plate of CrFK cells to be 75-80% confluent on Lab 5, Day 2/Lab 6, Day 1 – Monday, March 7.] BIOL 513 – Lab 5: Virus characterization – Part 3. Thermostability and pH sensitivity 80 LAB 5 INTRODUCTION TO VIRUS CHARACTERIZATION Thermostability and pH Sensitivity Viruses vary widely in their response to heat. Some viruses are quickly inactivated at higher temperatures, whereas others are much more heat stable. Knowledge of the thermostability of a given virus can predict if a given procedure will result in the destruction or preservation of viral activity....
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Lab5-2011 - BIOL 513 – Lab 5 Virus characterization –...

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