User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2011/11/02

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Objective

To synthesize gold nanoparticles, to perform DNA transformation on the PCR product prepared the previous day, to make an LB agar plate, and to run a gel to determine if PCR was successful.

Description

Synthesis of Gold Nanoparticles

  1. 1 mL of BSA, 7 mL of water, and 2 mL of HAuCl4 were placed in a reaction tube, in this order.
  2. The test tube was placed in an oven set to 80°C for 150 minutes. Every 30 minutes, the mixture was taken out of the oven for 10 minutes.

LB Agar Plate

  1. 0.875 g LB, 0.7 g Agar, and 35 mL of H2O were mixed in an Erlenmeyer flask.
  2. The contents of the flask were then autoclaved
  3. Before the agar completely cooled, 35μL of ampicillin was added to the flask.
  4. The contents of the flask were then poured onto a petri dish and allowed to solidify.

DNA Transformation

  1. Non-methylated DNA was digested with 1μL of Dpnl.
  2. Sterile reaction tube and DNA were placed in an ice bucket for 15 minutes.
  3. 5 μL of DNA and 30 μL of cells were added to the bottom of the tube.
  4. The tube was then incubated on ice for 30 minutes.
  5. The DNA and cells were heat shocked at 42°C for 30 seconds using the heat block.
  6. The reaction tube was incubated on ice for 5 minutes.
  7. 250 μL of SOC media was added to the reaction tube.
  8. The reaction tube was then incubated in the shaker for one hour at 37°C.
  9. 100 uL of the cell/DNA mixture in the reaction tube was spread on the LB agar plate.


Data

  • Colonies did not grow on the LB agar plate
  • The reaction mixture for nanoparticle synthesis produced a brown clump (brain) in a clear solution.

Notes

This area is for any observations or conclusions that you would like to note.


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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