Lab2-2011

Lab2-2011 - BIOL 513 Lab 2: Introduction to virus...

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Unformatted text preview: BIOL 513 Lab 2: Introduction to virus quantification 27 Lab 2: Introduction to virus quantification- lab designed for 12 groups of 2 students Equipment for Lab 2: Day 1 Day 2 Day 3 Day 4 Day 5 CO 2 incubator + + + + Light microscope - + - + Inverted phase contrast microscope + + + - 6-ft biosafety cabinet 2 - - 2 Hemacytometer with coverslip - 12 - 12 Hand counter - 12 - 12 Vortex 6 - 6 - 2-liter vacuum flask with aspirator 2 2 2 2 Pipetman, 40-200- L 12 12 12 12 8-channel Pipetman, 50-300- L 6 6 - 6 Safety bulb 12 12 12 12 250-mL beaker 12 12 12 12 Rack for microcentrifuge tubes - 12 - 12 Rack for 12 75-mm tubes 12 - 12 - Rack for 15-mL tubes - - 12 - Rack for 50-mL tubes 12 12 - 12 Supplies & Consumables for Lab 2: Day 1 Day 2 Day 3 Day 4 Day 5 96-well tissue culture plate cluster - - - 24 12-well plate cluster - 12 - - Microcentrufuge tube - 12 - 12 Sterile 12 75-mm tube with snap cap 168 - 120 - 50mL sterile conical tube - - - - 1-mL pipet 24 - 12 - 5-mL pipet 24 - - - 10-mL pipet - 27 - 36 25-mL pipet - - 12 - Sterile Pasteur pipet 24 6 36 6 Sterile dilution trays 24 18 - 12 Sterile filtered pipet tips in trays 12 12 12 12 Yellow pipet tips in trays - 12 - - KimWipes + + - + Paper towels - - - + Crystal violet stain in 50% methanol - + - + 0.4% Trypan blue stain - + - + 70% ethanol in spray bottle 6 6 6 6 Ethanol - + - - 1% methylcellulose overlay - - 12 - BIOL 513 Lab 2: Introduction to virus quantification 28 Lab 2: Introduction to virus quantification - lab designed for 12 groups of 2 students Materials for Lab 2 provided by the instructor: Day 1: Tuesday, February 1, 2011 FCV, 100 L at 5x10 5 PFU/mL in 12 75-mm snap cap tube 12 FHV, 100 L at 5x10 5 PFU/mL in 12 75-mm snap cap tube 12 CrFk growth medium (DMEM +10% FCS, 1 AA), 30 mL in 50-mL tube 12 Extra 96-well plate clusters with seeded cells 6 Day 2: Monday, February 7, 2011 CrFk cell suspension, freshly trypsinized, 25 mL @ 2-2.5 10 5 cell/mL 12 (from 3-4 150-mm plates @ 1.8 10 7 cell/plate to get total 60-120 10 6 cells) Extra CrFk medium Day 3: Tuesday, February 8, 2011 FCV, 100 L at 5x10 5 PFU/mL in 12 75-mm snap cap tube 12 FHV, 100 L at 5x10 5 PFU/mL in 12 75-mm snap cap tube 12 Viral dilution and inoculation medium (M199+2%FCS, 1 AA), 14 mL in 15-mL tube 12 Extra 12-well plate clusters with seeded cells 6 Day 4: Monday, February 14, 2011 CrFk cell suspension, freshly trypsinized, 25 mL @ 4 10 5 cell/mL 12 (from 5-6 150- mm plates @ ~2 10 7 cell/plate to get total 120 10 6 cells) Extra CrFk medium BIOL 513 Lab 2: Introduction to virus quantification 29 LAB 2 INTRODUCTION TO VIRUS QUANTIFICATION Infectivity, or the ability to invade and parasitize cells in order to multiply, is the most...
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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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Lab2-2011 - BIOL 513 Lab 2: Introduction to virus...

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