Katherine K. Jacobs: Notebook

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(New page: 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...)
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Revision as of 17:44, 6 February 2008

Lab Introduction January 30, 2008:=

Contents

=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
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