- Synthesis of Gold Nanoparticles using 0.25 mM HAuCl4 and 15 uM BSA in 50 mM Tris Buffer (pH 7.55).
- Transform DNA from experiment performed on 20th of September into E. Coli cells and then plate the cells on LB/Agar.
Synthesis of BSA Conjugate Gold Nanoparticles
- Approximately 0.581 mL of HAuCl4 (0.25mM), 0.974 mL BSA (15uM),and 8.45 mL Tris Buffer pH 7.55 (50 mM) were placed in a test tube.
- The small amount of the mixture was pipetted into a cuvette.
- The test tube was placed into an oven set at 80°C.
- A parafilm was used to cover the cuvette, which contains the sample, and the cuvette was placed into a UV-Vis Spectrophotometer (Shimadzu 2550).
- The temperature of the UV-Vis was set at 80°C and a spectra of the sample, which was kept inside the UV-Vis, was taken every 30 minutes for two hours.
Preparation of Agar Plate
- Approximately 5g of liquid broth was added to 4g of Agar in 200ml of H2O.
- The mixture was heated for 3 minutes in microwave.
- Approximately 200ul of 100mg/ml of Ampicilin was added to mixture after cooling to a temperature less than 60°C.
C. DNA Transformation
- Approximately 1 uL DpNI Digest was added to the DNA sample.
- The sample was placed into a thermocycler for 1 hour at a temperature set at 37°C.
- DNA was placed into an ice bath for 15 minutes.
- Approximately 5ul of the DNA sample and 50ul of Nova Blue Cells were placed into an eppendorf tube and were mixed.
- The mixture was placed on ice for 30 minutes.
- The mixture was heat shocked for 30 seconds and was then placed on ice for 5 minutes.
- Approximately 250ul of SOC media was pipetted into the mixture and the entire mixture was placed into a shaker for 1 hour at 37 degrees Celsius.
- Approximately 200ul of the mixture was pipetted into the plate and was spread.
- The plate was placed into a 37°C oven in an inverted position and was stored overnight.
- The UV-Vis Spectrum was taken at the following times.
T0 - 1:45
T1 - 2:15
T2 - 2:45
T3 - 3:15
T4 - 3:45
- The graph below is the UV-Vis Spectra of HAuCl4/BSA in Tris buffer (pH=7.5) obtained in the wavelength range of 200-800nm.
- The graph below exhibits the change in absorbance at 550nm over a period of 2 hours.
- According to the two graphs, the change in Absorbance was small.
- According to observations, the solution was clear and no fibers formed.
- This indicated that no conjugated gold nanoparticles formed.
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