Lab8-2011

Lab8-2011 - BIOL 513 Lab 8. Virus concentration Lab 8 Virus...

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BIOL 513 – Lab 8. Virus concentration 125 Lab 8 – Virus concentration - lab designed for 12 groups of 2 students Equipment for Lab 8: Day 1 Day 2 Day 3 Day 4 Day 5 CO 2 incubator - + + - 6-ft biosafety cabinet - + - - -80°C freezer + - - - Refrigerator + + - - Light microscope - + - - Inverted phase contrast microscope + + + - Hemacytometer with coverslip - 12 - - Hand counter - 12 - - Pipetman, 40-200- µ L - 12 12 - 8-channel Pipetman, 50-300- µ L - 6 - 6 2-liter vacuum flask with aspirator 2 - 2 2 Vortex 6 6 6 - Safety bulb 12 12 12 3 250-mL beaker - - 12 - Dialysis clamps, medium size - 24 - - Forceps - 12 - - Rack for microcentrifuge tubes 12 12 24 - Rack for 12 × 75-mm tubes - - 12 - Rack for 15-mL tubes 12 12 - - Rack for 50-mL tubes - - 24 - Supplies & Consumables for Lab 8: Day 1 Day 2 Day 3 Day 4 Day 5 96-well tissue culture plate - 12 - - Microcentrifuge tube 12 24 - - Sterile 12 × 75-mm tube with snap cap - - 202 - 15-mL conical tube 12 12 - - 1-mL pipet 12 12 36 - 10-mL pipet 24 48 - 2 Sterile Pasteur pipet 12 - 12 6 Sterile dilution trays - 12 - 12 Yellow pipet tips in trays - - - 12 Sterile filtered pipet tips in trays - 12 12 - PEG 8000, 2 g in a 15-mL conical tube 12 - - - PEG 20,000, 10 g in a 60-mm dish - 12 - - Dialysis membrane * Distlilled water in wash bottle - 12 12 - 0.4% Trypan blue stain - + - - Crystal violet stain - - - + 70% ethanol in spray bottle 6 6 6 6 Paper towels - - - + KimWipes - + + - Also check the corresponding section of Lab 9, which begins on day 4 of Lab 8.
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BIOL 513 – Lab 8. Virus concentration 126 Lab 8– Virus concentration - lab designed for 12 groups of 2 students Materials for Lab 8 provided by the instructor: Day 1, Tuesday, April 5, 2011 PBS, 1 mL in 1.5 mL tube – 12 Instructors: You will need to plate CrFk cells in T-75 flasks on Friday, April 8 and infect the flasks with FHV on Saturday, April 12. Day 2, Monday, April 11, 2011 12 mL suspension of freshly trypsinized CrFk cells @ 4 × 10 5 cell/mL – 12 (from 2-3 150 mm plates @ ~2 × 10 7 cells/plate to get total 60 × 10 6 cells) FHV virus suspension, 10 mL in 75 cm 2 flask & 3 dpi – 12 Dialysis membrane, 10-12K MWCO, pre-soaked, ~5 cm × 15 cm flattened tube – 12 Extra CrFk medium Day 3, Tuesday, April 12, 2011 CrFk growth medium, 40 mL in 50 mL conical tube – 12 (for 12 groups) PBS, 1 mL in 1.5 mL tube – 12 Day 4, Monday, April 18, 2011 None Also check the corresponding section of Lab 9, which begins on day 4 of Lab 8.
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BIOL 513 – Lab 8. Virus concentration 127 LAB 8 VIRUS CONCENTRATION For certain assays it can be useful to purify and concentrate viruses collected from either biological fluids or cell culture supernates. Purifying virus that is grown in tissue culture is relatively simple, as the cell culture media used are relatively free of contaminants and unwanted
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This note was uploaded on 01/22/2011 for the course BIO 513 taught by Professor David during the Spring '10 term at Kansas.

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Lab8-2011 - BIOL 513 Lab 8. Virus concentration Lab 8 Virus...

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