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

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==Objective==
==Objective==
*Conjugated Gold Nanoparticle Synthesis.  
*Perform Conjugated Gold Nanoparticle Synthesis using the same amount of BSA and HAuCl<sub>4</sub> used on November 1<sup>st</sup>, 2011.  
*Transform the mutated DNA that was produced using PCR on November 1<sup>st</sup>, 2011.
*Transform the mutated DNA that was produced using PCR on November 1<sup>st</sup>, 2011.


Revision as of 01:01, 9 December 2011

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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.

Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized. .


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