User:Daniel-Mario Larco/Notebook/AU Photosynthesis Lab/2014/06/05

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Today we will be purifying our protein solution


  1. The dialysis tubing containing the protein was removed form the Tris buffer ph 9
  2. The content of the tube was poured into a 50mL falcon tube
  3. A filtration system was setup on a vacuum using membrane filter
  4. The collection tube was placed on ice during the filtration
  5. The protein solution was poured in in 10mL increments
  6. The solution was filtered a second time
  7. After a second round of filtration, the solution was placed in dialysis tubing
  8. The tube containing our protein solution were placed in 50mM Tris buffer at pH 9 in a cold room overnight


During this lab session, solutions LB Broth were prepared. A solution of Ampicilin was prepared to be added to the broth and agar.


1. The correct concentration for dissolving LB Broth Media into water is 25g per liter of water.

- For the 1L LB Broth solution, 25g of LB Broth Media were dissolved into 1L of water

- For the 25mL LB Broth solution, 0.625g of LB Broth Media were dissolved into 25mL of water

The solution are placed in the autoclave at 121 degrees C for an hour.

2. In order to prepare the ampicilin solution, 500mg of Ampicilin Sodium Salt were dissolved into 5mL of sterilized water. This solution was prepared in a sterilized falcon tube using sterile equipment.

3. Once the solutions cool, 1mL of the ampicilin solution is added to the 1L flasks and 25uL were added to the 25mL flasks

4. Cells were transferred to the 25mL flasks and left to spin overnight

    1. K31cAsc Hb BL21DE5 cells