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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 mL of BSA, 7 mL of water, and 2 mL of HAuCl4 were placed in a reaction tube, in this order.
- 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
- 0.875 g LB, 0.7 g Agar, and 35 mL of H2O were mixed in an Erlenmeyer flask.
- The contents of the flask were then autoclaved
- Before the agar completely cooled, 35μL of ampicillin was added to the flask.
- The contents of the flask were then poured onto a petri dish and allowed to solidify.
DNA Transformation
- Non-methylated DNA was digested with 1μL of Dpnl.
- Sterile reaction tube and DNA were placed in an ice bucket for 15 minutes.
- 5 μL of DNA and 30 μL of cells were added to the bottom of the tube.
- The tube was then incubated on ice for 30 minutes.
- The DNA and cells were heat shocked at 42°C for 30 seconds using the heat block.
- The reaction tube was incubated on ice for 5 minutes.
- 250 μL of SOC media was added to the reaction tube.
- The reaction tube was then incubated in the shaker for one hour at 37°C.
- 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.
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Experimental Biological Chemistry
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