Afraid i was gonna leave without giving you a goodbye

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__________________________________________ “Afraid I was gonna leave without giving you a goodbye kiss?” ‘I'd just as soon kiss a Wookie.’ “I can arrange that. You could use a good kiss.” © 1980 LucasFilm Ltd.
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Lab 2 Page 9 Growth Curve Sample Readings Work together as a Team when taking Samples and OD Reading. It’s probably best if one of you does one particular Aspect (say, taking the Samples) and the other does another Aspect (say, taking and recording the Spec Readings). 1. At each Sample Timepoint retrieve your Flask of inoculated Growth Medium from the Water Bath and carry it back to your Bench. • Be sure the Contents have been thoroughly mixed by swirling 2. Use a P1000 Microtiter Pipette to measure 1000 μ l (1 ml) of your Growth Medium into your Cuvette. 3. Return your Flask to the Water Bath. Quick like a Bunny get that Flask back into the Shaking Water Bath so your E. coli will remain at 37°C. If your Flask is out of the Water Bath for longer than about a Minute at every Sample Timepoint, the Temperature will Porpoise and the Bacteria will not be able to maintain Exponential Growth. The whole Point of this Exercise is to maintain Exponential Growth, so -- Quick like a Bunny -- get that Flask back into the Water Bath 4. Insert your Cuvette into the Sample Holder. 5. Read the Absorbance from the LCD Screen. Unless you ʼ re very, very Good don ʼ t try and plot your OD Data on Semi-Log Graph Paper until after you ʼ ve taken 3 or 4 Readings. 6. Remove your Cuvette and decant the Contents into the Waste Beaker. 7. Rinse your Cuvette using your DI Water Bottle. Decant each of these Rinses into the Waste Beaker (they contain Butt-loads of Bacteria). 8. After the Final Rinse (say, the third Rinse) use a Strip of Paper Towel to blot any Excess Rinse from the Rim of your inverted Sample Tube. • Remember, this Paper Towel is now Contaminated Waste. • One Paper Towel torn into Strips should get you through the entire Growth Curve Exercise. 9. Invert your Cuvette over a Strip of Paper Towel until the next Reading. ________ “Let go.” ‘Shh.’ “Let go, please.” ‘Don't get excited.’ “Captain, being held by you isn't quite enough to get me excited.” ‘Sorry sweetheart. I haven't got time for anything else.’ © 1980 LucasFilm Ltd.
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Lab 2 Page 10 Graphing your Growth Curve Data 1. Record your Optical Density (OD) [Absorbance] Readings in your Lab Notebook. • Be sure and accurately record the Time for each Sample 2. Wait until you have three or four (3 or 4) OD Readings before you begin to transfer your Readings from your Lab Notebook to a Sheet of Semi-Log Graph Paper. 3. Once you’ve got those OD Readings determine the Values for your Decades on your Semi-Log Graph Paper by doing some quick “What if?” Plots. When all else fails, Think! Your First OD Reading was probably very, very Low. That Tube looked almost Clear and so you knew it would be close to “0” (or close to 100% Transmission, which sometimes is easier to Visualize).
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