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2011 Week 2 Lab - BIO 122 FINAL

2011 Week 2 Lab - BIO 122 FINAL - Stanford and Kabnick Fall...

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Stanford and Kabnick Fall 2011 Bio 122: Week 2 Lab Goals : 1. Learn how to use a micropipettor 2. Use a blender and centrifuge to isolate your cell fractions for the SDH assays 3. Aliquot your mitochondrial and crude cell fractions for use over the next two weeks 4. Begin to develop a hypothesis relating your assigned reagent to its effect on SDH activity Preparing for Lab : Review the information on the proper use of a micropipettor Review the protocol on cell fractionation Notes :
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Stanford and Kabnick Fall 2011 Use of Micropipettors To transfer small volumes of liquid, you will use micropipettors. This exercise will help you to become familiar with using micropipettors so that you can use them effectively in this and other subsequent lab periods. Here are some general rules for operation: 1. Set the volume by turning the volume adjustment knob until you see the correct volume on the indicator. Never rotate the volume adjustment knob beyond the range of the micropipettor . 2. Attach a disposable tip to the narrowed end of the micropipettor without touching it with your fingers. Make sure the tip is pressed on firmly to ensure a tight seal. Never use micropipettors without tips . P-10 models use white tips (0.5-10 µl) P-20, P-100 and P-200 models use yellow tips (10-100 µl) P-1000 models use blue tips (100-1000 µl) 3. Micropipettors have two “stops.” Depress the plunger to the first place you feel a catch. This is the first stop. You will find that with a bit more applied pressure, you can depress the plunger to a second “stop.” Practice finding the first and second stops before you attempt to draw liquid into your tip. This will allow you to feel comfortable using your micropipettor. 4. Before putting your tip into the liquid, depress the plunger to the first stop. This stop is the stop that is calibrated to the volume on the indicator. 5. Hold the micropipettor vertically and place the tip in the liquid to be pipetted. Do not immerse the tip completely . The tip should be inserted into the liquid so that only the bottom of the tip is immersed. You do not want to immerse the pipettor itself into the liquid for several reasons: 1) you do not want to get liquid into the mechanics of the pipettor, as it may damage the pipettor.
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2011 Week 2 Lab - BIO 122 FINAL - Stanford and Kabnick Fall...

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