Lab7-2011

Lab7-2011 - BIOL 513 Lab 7. Viral Replication - One-step...

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BIOL 513 – Lab 7. Viral Replication - One-step growth curve 105 Lab 7: One-step growth curve - lab designed for 12 groups of 2 students Equipment for Lab 7: Day 1 Day 2 Day 3 Day 4 Day 5 CO 2 incubator + + + + 6-ft biosafety cabinet - + - - -80°C freezer + + - - Light microscope - + - - Inverted phase contrast microscope + - + + Bench-top microcentrifuge + - + - Hemacytometer with coverslip - 12 - - Hand counter - 12 - - 2-liter vacuum flask with aspirator 2 - 2 2 Pipetman, 40-200- µ L 12 12 24 - 8-channel Pipetman, 50-300- µ L - 6 - 6 Pipetman, 100-1000- µ L 12 - - - 250-mL beaker - - - 12 Safety bulb 12 12 24 12 Forceps - 12 - - Vortex - 6 6 - Rack for microcentrifuge tubes 12 12 24 - Rack for 12 × 75-mm tubes - - 48 12 Rack for 15-mL tubes 12 12 - 12 Rack for 50-mL tubes 12 12 24 - 37°C water bath - + - - Supplies & Consumables for Lab 7: Day 1 Day 2 Day 3 Day 4 Day 5 75-cm 2 flask - 12 - - 96-well tissue culture plate - 48 - - 1-mL pipet 12 - 24 - 5-mL pipet - 12 - 12 10-mL pipet 36 36 - 26 Sterile Pasteur pipet 6 - 48 6 Microcentrifuge tube 192 12 - - 12 × 75-mm sterile tube with snap cap 12 - 1536 - 125-250-mL sterile bottle - - 12 (for the instructor) Sterile dilution trays - 12 - 12 Yellow pipet tips in trays 12 - 24 12 Sterile filtered tips in trays - 12 - - Blue pipet tips in trays 12 - - - 0.4% Trypan blue stain - + - - Crystal violet stain - - - + 70% ethanol in spray bottle 6 6 6 6 Paper towels - - - + KimWipes - + - +
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BIOL 513 – Lab 7. Viral Replication - One-step growth curve 106 Lab 7 – One-step growth curve - lab designed for 12 groups of 2 students Materials for Lab 7 provided by the instructor: Day 1, Tuesday, March 15, 2011 (Note to students and instructors: on this day, students will need to come back to the lab several hours after the lab period has finished, including after hours. Be sure to make the proper arrangements.) FCV in M199+2%FCS, 1 mL @ 2 × 10 7 pfu/mL – 12 M199+2%FCS, 2 mL in 12 × 75-mm tube – 12 PBS, 25 mL in a 50-mL tube – 12 CrFk growth medium, 10 mL in a 15-mL tube – 12 12-well plate with CrFk cells in 8 wells – 12 Extra PBS, M199+2%FCS, CrFk growth medium Day 2, Monday, March 28, 2011 50 mL suspension of freshly trypsinized CrFk cells @ 4 × 10 5 cell/mL – 12 (from 10 150- mm plates @ ~2 × 10 7 cells/plate to get total 200 × 10 6 cells) CrFk growth medium (DMEM +10% FCS, 1X AA), 10 mL in a 15-mL conical tube – 12 Extra CrFk medium Day 3, Tuesday, March 29, 2011 CrFk cell culture medium (DMEM +10% FCS, 1 × AA), 120 mL in a suitable sterile flask – 12 Extra 96-well plate with seeded cells – 6 Extra CrFk medium Day 4, Monday, April 4, 2011 FCV in M199+2%FCS, 2 mL @ 5 × 10 5 pfu/mL in a 12 × 75-mm tube with snap-cap – 12 CrFk growth medium (DMEM +10% FCS, 1 × AA), 10 mL in a 15-mL conical tube – 12
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BIOL 513 – Lab 7. Viral Replication -
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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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Lab7-2011 - BIOL 513 Lab 7. Viral Replication - One-step...

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