Objective
AuNP VIII
To synthesize AuNP 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, 09/07/11, 09/27/11, 09/28/11, 10/19/11, 10/26/11, and 11/01/11.
DNA Transformation II
To transform DNA coding for mutant GFP (mutated on 09/20/11) into Novablue cells.
In addition, to repeat the experiment performed on 09/27/11.
Gel II
To determine whether or not the PCR performed on 11/01/11 of the DNA coding for a mutant GFP was successful using agarose gel electrophoresis.
In addition, to repeat the experiment performed on 09/21/11.
Description
AuNP VIII
- Combine 8 mL of water, 1 mL HAuCl4 (2.9 mM), and 1 mL BSA (1.55 μM)in that order a test tube.
- Place test tubes in an oven at 80°C for 30 min.
- Remove test tubes from oven for 10 min.
- Repeat steps 2 and 3 for 3.5 hr.
DNA Transformation II
- Digest non-methylated DNA with 1 μL Dpnl.
- Make LB/agar plates
- Mix 0.875 g LB, 0.7 g agar, and 35 mL of water.
- Autoclave.
- Add 35 μL ampicillin before agar completely cools.
- Place sterile tube and DNA in ice bucket for 15 min.
- Add 5 μL of DNA and 40 μL 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.
Gel II
- Make a 1% agarose gel by adding 0.25 g of 0.01 g/mL of agarose to 25 mL of 1x tris base, acetic acid, and ethylenediaminetetraacetic acid (TAE) buffer and heating the mixture in the microwave for 40 s.
- Mix 2 μL of 6x gel loading dye with 10 μL of the DNA sample.
- Prepare a ladder containing DNA, glycerol, and gel loading dye.
- Load DNA solutions and run the gel at 80 V for 40 min.
- Remove gel and place in TAE and ethidium bromide (EtBr) solution, mix for 15 min.
- Rinse gel with TAE for 5 min.
Data
Notes
Order of DNA gel:
Ladder|Tamra 1|Tamra 2|Group 1|Group 2|Group 3|Group 4|Group 5
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