User:Nader A. Khamis/Notebook/Experimental Biological Chemistry AU Fall 2011/2011/11/02

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Objective

  • Perform Conjugated Gold Nanoparticle Synthesis using the same amount of BSA and HAuCl4 used on November 1st, 2011.
  • Transform the mutated DNA that was produced using PCR on November 1st, 2011.

Description

Conjugated Gold Nanoparticle Synthesis:

  1. Approximately 1ml of BSA stock solution (15.5µM), 1ml of HAuCl4 stock solution (2.9mM) were pipetted into a test tube and was filled with 8ml of H2O.
  2. The following steps were then executed:
  • Leave solution in oven for 30 minutes.
  • Remove solutions from oven and keep them in room temperature for 10 minutes.
  • Place solutions back in oven.
  • Repeat cycle for 2 hours.

DNA transformation

The LB/agar plate:

  1. 0.875g LB + 0.7g Agar + 35ml H2O were mixed.
  2. The solution was autoclaved for 1hours and a half
  3. Approximately 35µL of Antibiotics (Ampicillin) and 35mL of water were added.
  4. The solution was added into a plastic Petri dish until the surface of the plate was completely covered.
  5. The solution was left to solidify.

Prepare the cells:

  1. Place the eppendorf tube containing DNA solution from PCR into the ice.
  2. Approximately 5µL of DNA solution was pipetted to a solution containing 30µL of cells.
  3. The cells were incubated on ice for 30 minutes
  4. Following 30 minutes, the solution was placed 42°C heatblock for 30 seconds.
  5. Incubate in ice for 5 minutes
  6. Approximately 250μL of SOC media was pipetted into the cell solution.
  7. The cell solution was incubated in shaker at 37°C for 1 hour.

Data

  • For the BSA/HAuCl4 solution, purple fibers formed .

Notes

The molar ratio of HAuCl4 to BSA was 187.

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