Lab1-2011

Lab1-2011 - BIOL 513 Lab 1 Introduction to cell culture Lab...

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BIOL 513 – Lab 1: Introduction to cell culture 1 Lab 1: Introduction to cell culture – lab designed for 12 groups of 2 students. Equipment for Lab 1: Day 1 Day 2 Day 3 Day 4 Day 5 CO 2 incubator + + + Inverted phase contrast microscope + + + Light microscope + + + Hemocytometer with coverslip - 12 12 Hand counter - 12 12 Pipetman, 40-200- µ L - 12 12 8-channel pipetman, 20-200- µ L - - 6 Safety bulb - 12 12 250-mL beaker - 12 12 Rack for microcentrifuge tubes - 12 12 Rack for 12x75-mm tubes 12 12 12 Rack for 15-mL tubes - - 12 Rack for 50-mL tubes - 12 12 Forceps 12 - 12 37°C water bath - + + Supplies & Consumables for Lab 1: Day 1 Day 2 Day 3 Day 4 Day 5 25-cm 2 flask - 24 - 96-well plate cluster - - 12 Sterile dilution tray - - 12 12x75-mm test tube - 36 12 15-mL conical sterile tube - - 12 Microcentrufuge tube - 12 12 1-mL pipet - 60 48 5-mL pipet - 108 96 Filtered sterile 200- µ L pipet tip trays - - 12 Staining jars (Petri dishes) 12 - - Microscope slides 12 - - pH paper with color chart 12 - - Mounting medium + - - Yellow pipet tips - + - Wright-Giemsa stain + - - 0.4% Trypan blue stain - + + 70% ethanol in spray bottle - 12 12 Wood applicator sticks + - - Toothpicks + - - KimWipes - + + Distilled water + + +
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BIOL 513 – Lab 1: Introduction to cell culture 2 Lab 1: Introduction to cell culture – lab designed for 12 groups of 2 students. Materials for Lab 1 provided by the instructor: Day 1: Monday, January 24, 2011 25-cm 2 flasks of different cell lines o CrFk (Crandall feline kidney) cell line @ ~50% confluence – 12 o Vero (African green monkey kidney) cell line @ ~50% confluence – 4 o NB-4 (human promyelocyte) line @ 5x10 6 cells/mL – 12 Coverslips with cells: o CrFk @ 30-50% confluence – 12 o Vero @ 30-50% confluence – 12 Sheathed 18 gauge needles with curved tip – 12 12x75-mm tubes with cell culture media at pH 6.5, 7.4 and 8.0 – 12 each Day 2: Tuesday, January 25, 2011 25-cm 2 flasks of different cell lines o CrFk (Crandall feline kidney) cell line @ ~50% confluence – 12 o NB-4 (human promyelocyte) line @ 5x10 6 cells/mL – 12 15-mL tube with 6 mL Dulbecco’s-Phosphate Buffered Saline w/o Ca 2+ and Mg 2+ (D- PBS) – 12 12x75-mm tube with 2 mL 0.05% trypsin-EDTA – 12 CrFk growth medium (DMEM +10% FCS, 1x AA), 10 mL in 15-mL conical tube – 12 NB-4 growth medium (RPMI +10% FCS, 1x AA), 10 mL in 15-mL conical tube – 12 Day 3: Monday, January 31, 2011 150 mm dish of confluent CrFk in case of bacterial contamination – 1 (3.75x10 7 cells, enough to seed 12 96-well plate clusters) Dulbecco’s-Phosphate Buffered Saline w/o Ca 2+ and Mg 2+ (D-PBS), 15-mL tube – 12 0.05% trypsin-EDTA, 12x75-mm tube with – 12 CrFk cell culture medium (DMEM +10% FCS, 1x AA), 20 mL in 50-mL tube – 12 Extra CrFk medium, D-PBS, trypsin-EDTA To be re-used next Tuesday These remain from Monday
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This note was uploaded on 01/22/2011 for the course BIO 518 taught by Professor Heft during the Spring '10 term at Kansas.

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Lab1-2011 - BIOL 513 Lab 1 Introduction to cell culture Lab...

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