Objective
To synthesize gold nanoparticles (Au NP) using the procedure developed here:
Bakshi, Mandeep S.; Kaur, Harpreet; Khullar, Poonam, Banipal, Tarlok, S.; Kaur, Gurinder; Singh, Narpinder Journal of Physical Chemistry C 2011, 115(7), 2982-2992
http://pubs.acs.org/doi/abs/10.1021/jp110296y
In addition, to repeat the experiment performed on 08/31/11 and 09/07/11.
Description
AuNP III
- Combine 10 mL of 50 mM acetate buffer (pH 5.46) with 1.1×10-3 g of .0015 mM bovine serum albumin (BSA) and 4.0×10-4 g of 0.25 mM chloroauric acid (HAuCl4) in a capped test tube at room temperature.
- Repeat with 10 mL of distilled water instead of acetate buffer.
- Take UV vis spectra (200 nm to 800 nm) of buffer, water, and both reaction mixtures.
- Place reaction mixtures in an oven at 80°C for 6 hr under thermostatic conditions.
- Continue to take UV vis spectra of reaction mixture every 30 minutes.
- After 2.5 hr, remove from oven and cool to room temperature; leave sitting overnight.
Protein Expression
Starter Culture Media, Protein Expression
- Make expression culture media.
- Prepare LB in a 250 mL Erlenmeyer flask.
- 0.875 g of LB
- 35 mL of water
- Cover the flask with foil.
- Autoclave.
- Allow flask to cool.
- Add 35 μL of 1000X ampicillin.
- Inoculate with the bacteria you are culturing using one of the two following methods:
- Using sterile methods, retrieve a single colony from a petrie dish and drop it into the flask
- Using sterile methods, add 5uL of a glycerol stock to the flask
- Place the flask in a shaker/incubator and culture overnight at 37°C and 200 rpm.
DNA Transformation
- Digest non-methylated DNA with 1 μL Dpnl.
- Make LB/agar plates with ampicillin.
- Place sterile tube and DNA in ice bucket for 15 min.
- Add 5 μL of DNA and 40μL of cells to the bottom of the tube.
- Incubate on ice for 30 min.
- Heat shock DNA/cells at 42°C for 30 s.
- Incubate on ice for 5 min.
- Add 250 μL of SOC media.
- Incubate in shaker at 37°C for 1 hr.
- Spread 100 μL of cells on LB/agar plate.
- Store plate (inverted) in 37°C oven overnight.
Data
Notes
AuNP solution turned purple while in the oven.
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