User:Susan Schultz/Notebook/Experimental Biological Chemistry/2011/11/02
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Run PCR Product on Gel
Transform PCR Product into E. Coli
Run additional AuNP synthesis reaction with fresh gold stock
Reaction 1- 1mL 15.5μM BSA + 1mL 2.5mM HAuCl4 removing from heat every 30 minutes for 10 minutes
Reaction 2- 1mL 15.5μM BSA + 1mL 2.5mM HAuCl4 heating continuously (some fluctuation from opening oven door)
Remove wax from PCR Product
Add 1μL of DpnI and heat in heat block at 37°C for one hour to digest non-methylated DNA
Prepare 1.2% Agarose Gel: Add 0.25 g of 0.01 g/mL of agarose, 25 mL of 1x tris base, acetic acid, and TAE buffer in flask. Microwave for 40 seconds.
Pour gel and allow to solidify
Transfer 10μL of the PCR product to second PCR and add 2μL of 6X loading buffer
Load solution into gel, along with ladder
Prepare agar plate: 0.875 g LB, 0.7 g agar, and 35 mL of water.
As mixture cools add 35 μL ampicillin.
Add 5μL of PCR product to 30μL NovaBlue Competent E.coli in sterile PCR tube
This area is for any observations or conclusions that you would like to note.