User:Noy Kaufman/Notebook/EBC I AuNP/2017/09/13
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1.Electrophoresis Cell was assembled, comb and tape were removed from the gels and the wells were rinsed with running buffer. Inner and outer buffer chambers were rinsed with running buffer 2. Samples preparation: - microcentrifuge tubes used to prepare solutions below 10uL of BSA stock was diluted to 1mL with the Glycine-HCl buffer prepared 9/12 10uL of this diluted sample was then mixed with 10uL of the SDS-PAGE running buffer This was repeated for arginine solution, AuNP from BSA solution and AuNP from your arginine. -100 μL of 0.1 wt% poly(l-lysine) from fridge was dilute to 1mL with the Glycine-HCl buffer Samples were heated for 5 minutes at 95C (in the thermocycler. 20uL of protein ladder was loaded into gel 20uL of prepared samples were loaded into the appropriate lane of your gel 3. Performed electrophoresis Ran 30 min at 200 V, 0.05 A, and 10 W While experiment was running,solutions below were prepared: a. Stain for gel: Prepared 100 mL of each solution Placed gel in Fixative Solution (40% methanol, 10% acetic acid, 50% water) for 30 minutes Placed gel in Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for 1 hour** Placed gel in Destain Solution (10% acetic acid, 90% water) for 15 minutes Repeated this step with fresh destain solution 2 more times
figure 1. solutions prepared and numbered
figure 2: gel layout
Figure 3. gel ladder
figure 4. gel results
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