Pre-Laboratory Preparation |
See "Materials List," page 11, for a complete listing of materials for this experiment.
Note:
Kit contains enough reagents for 10 gels
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Protein samples:
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Yellow Perch
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Walleye
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Chinook salmon
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High molecular weight protein marker
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2
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6-14 % SDS polyacrylamide gels
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2
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SDS running buffer packs 4X concentrate
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2
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Bottles, Ward's protein Stain (500 mL each)
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1
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Gel staining tray
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1
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Spatula (gel handler)
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5
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Plastic transfer pipets (6 mL)
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24
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Microcentrifuge tubes
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5
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Sheets of graph paper
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1
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"Student Study Sheet" Copymaster
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1
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"Student Analysis Sheet" Copymaster
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1
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Teacher's Manual
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(1)
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Prepare SDS running buffer (4 X) according to the following directions: To make concentrated stock solution, pour the contents of one packet into a 2-L beaker containing about 1-L of deionized or distilled water. Stir until the powder has dissolved completely. Fill beaker to the 1500 mL mark with additional distilled or deionized water. This is a 4X stock concentrate. To make 1-L of working buffer solution, just prior to use mix 750 mL of deionized or distilled water with 250 mL of buffer concentrate. One liter is enough to run one gel in a single tank and 2-L is enough to run two gels in a dual tank. NOTE: Although it is not usually necessary, filtration of running buffer helps assure crisp band imaging results. Store hydrated buffer concentrate and/or working buffer in a refrigerator.
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(2)
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Preparation of the lyophilized samples: To each lyophilized sample add 1 mL of distilled or deionized water. Dissolve the proteins completely by gently swirling the mixtures. Do not shake them. Store samples in a refrigerator. Shelf life of rehydrated samples is approximately one month.
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