Katherine K. Jacobs: Notebook: Difference between revisions
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=Lab Introduction January 30, 2008= | =Lab Introduction January 30, 2008= | ||
====3:30 PM Pipette Use==== | ====3:30 PM Pipette Use==== |
Revision as of 14:52, 6 February 2008
Lab Introduction January 30, 2008
3:30 PM Pipette Use
- Sterilization of the pipette with ethanol
- Pipette #1: 2 microliters blue solution and 5 microliters water
- Set pipette to 7 microliters to test accuracy of previous measurements
- Pipette #2: 20 microliters blue solution and 50 microliters water
Making Solutions:
- Solution #1: KCl Potassium Chloride 0.3 M
- Solution #2: MgCl2 Magnesium Chlroide 0.5 M
- Solution #3: NaCl Sodium Chloride 1 M
- Pour .5 of water amount into container, add salt then add the rest of the water to assist in mixture of two substances
- Preparation of Magnesium Chloride Solution:
(203.31 grams/ mole) x (0.5 moles/Liter) x (0.1 Liter) = 10.1655 grams/ 100 mL
- Preparation of Potassium Chloride Solution:
(74.56 grams/mole) x (0.3 moles/Liter) x (0.1 Liter) = 2.2368 grams/ 100 mL
4: 30 PM Autoclave:
- Mix 25 grams of LB mixture and 15 grams of Bactoagar to a 2 L flask and autoclave for 20 minutes
- Once autoclave was complete, the tape on the flask did not turn black indicating that the mixture was not successfully sterilized.
- Suggestion for solving problem – Autoclave for 40 minutes and set to Sterilize
5:30 PM Making Cell Cultures:
- LB with Ampicillin contains nutrients for the cells to grow
- Keep the culture tubes (with a loose lid) sterile by closing the bag after removing tubes
- The culture should get cloudy while the control should remain clear
- Sterilize the pipette with ethanol
- 2 mL cultures are optimal
- Fill both tubes with the LB first
- Sterilize the loop by placing in ethanol and then flaming
- When removing a colony from a plate, mark on the underside of the plate with a marker which colony has been taken
- Incute the culture overnight at 37 degrees C in a shaking water bath