User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2011/11/01

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Objective

To synthesize gold nanoparticles and to repeat the PCR reaction performed on 09/20/2011

Description

Gold Nanoparticle Synthesis

  1. Two reaction mixtures were prepared. Test tube 1 contained 1 mL of 3M HCl, 1 mL of 1.55μM BSA, and 8 mL of water, added in that order. Test tube 2 contained 1 mL of BSA, 7 mL of water, and 2 mL of 2.9μM HAuCl4, added in that order.
  2. The reaction tubes were placed in an oven set to 80°C for 3 hours. Ever 30 minutes, the tubes were taken out of the oven for 15 minutes.

PCR

  1. The PCR quick change manual was used to determine which forward and reverse primers should be used for the PCR. Prepared primers were used for the PCR reaction
  2. A reaction mixture was prepared with the following contents: 1 μL of template DNA, 1.25 μL of primer 1 (D1C F), 1.25 μL of primer 2 (D1C R), 5 μL of 10X Pfu reaction buffer, 1 μL of dNTP mix, 1 μL of PfuTurbo DNA polymerase, and 40 μL of water. The total volume of the PCR reaction mixture was about 50μL.
  3. 25 μL of Bio Rad Chill Out Wax was placed on top of the reaction mixture
  4. The reaction mixture was placed in the thermocycler for one cycle at 95°C for 30 seconds
  5. The reaction mixture was kept in the thermocylcer for 16 cycles with the profile shown below.
  6. The reaction mixture was left in the thermocycler for 24 hours at 4°C
       PCR Profile
   1) 95 °C for 30 seconds
   2) 55°C for 1 minute
   3) 68°C fr 3.6 minutes


Data

Gold Nanoparticle Synthesis Test tube 1 remained clear the entire reaction time. At the beginning of the reaction, test tube 2 had a yellowish color. At t=60, black clumps could be seen in solution. These black solid clumps remained for the majority of the rest of the reaction. When the reaction was placed in for an additional 30 minutes after reaching the 3 hour mark, the black solid clump changed to a dark red color.


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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