Performing the Experiment |
Materials: (per team of 5)
Samples in labelled microcentrifuge tubes:
Tube #1: Escherichia coli
Tube #2: Micrococcus luteus
Tube #3: Serratia marcescens DI
Tube #4: Unknown A
Tube #5: Unknown B
Tube #6: Molecular weight protein marker
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1
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10-20 % SDS polyacrylamide gel cassette *
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1
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Well forming comb
1 L (1X) SDS running buffer solution (for single tank)
Micropipettes (20 uL) |
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1
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Gel Staining tray
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1
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(non-sterile) Plastic transfer pipet (6 mL)
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1
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Sheet 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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Teacher's Note: A 13 lane gel cassette can be used by two student teams. Team #1 can load samples onto lanes 1-6 and Team #2 can load lanes 8-13.
Common Materials:
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WARD'S PAGE tank
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Ward's power supply
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1
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Bottle, Ward's protein stain (100 mL per gel)
Deionized or distilled water
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1
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Spatula "gel handler"
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WEAR EYE PROTECTION FOR ALL THESE STEPS
WARNING: The power supply produces high enough voltage to cause severe electrical shock if handled improperly. For safe operation of this unit, follow all directions and precautions. Assure that all students are thoroughly drilled in the procedure for using this unit, and making electrical connections, and that they are directly supervised by you. We strongly urge you to take the time to examine all components of your electrophoresis apparatus prior to each use. The maintenance inspection should include all cords, plugs, tank apparatus, and power supply.
Precautions:
Use the same precautions as you would with any other electrical device. This device produces high enough voltages (although low compared to general electrophoretic standards) to cause severe electrical shock.
Designate one area of the laboratory as an "electrophoresis area" where cells and power supplies are connected. Place the power supply near the tank to which it is to be connected but away from normal everyday activities.
Do not plug the power supply into a wall receptacle until the safety cover is positioned on the tank and all other electrical connections are properly made.
This unit uses a 3-wire grounded plug; for safety reasons it should not be used with a 2-wire receptacle with a conversion plug. Use a circuit tester to confirm proper polarity.
Do not operate in a damp or humid environment since any condensed moisture may short out electrical components. Assure that the power supply, electrical cord, and patch cords are clear from wetness or moisture.
Inspect all power cords, patch cords and banana jacks and plugs for any defects, such as cracked and dried out insulation and loose or wobbly banana jacks or plugs.
Do not come in personal contact, or allow metal or any conductive material to come in contact, with the reservoir buffer or the electrophoretic tank while the power supply is on.
The power supply may continue to produce some voltage even when the power has been turned off. To eliminate any risk associated with this event, follow all required steps given in the procedure. When disconnecting the electrophoretic chamber be sure the leads do not touch each other, come in contact with the buffer, or otherwise create any hazardous electrical condition.
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