Team:Arizona State/Lab/Protocols/Gel visualization
From 2011.igem.org
- Mix 1x TAE Buffer with 0.50g agarose in 250 mL Erlenmeyer flask. Swirl well.
- Microwave solution until it begins to boil. Gently swirl the flask and repeat the microwaving process until the agarose is completely dissolved and the solution is clear.
- Add 5 μL SYBR green and gently swirl the mixture. Let the solution sit for 5 minutes.
- Carefully pour the solution into the 50 mL gel tray and add in the desired comb. Let sit for 30-45 minutes until gel solidifies.
- Place gel tray in the -4 o C refrigerator for 10-15 minutes.
- Remove gel tray from refrigerator, remove rubber ends, and place the gel in electrophoresis chamber with the comb towards the negative end.
- Add in enough 1x TAE Buffer in the chamber to just barely create a thin film of buffer over the gel.
- Add 2 μL of each DNA sample to a strip of parafilm. Add 8 μL of loading buffer to each of those 2 μL samples.
- Add the necessary Hyperladder to the first well in the gel.
- Add each 10 μL mix of buffer and DNA into individual wells in the gel.
- Attach the electrophoresis chamber lid onto the chamber and plug into the power source.
- Set the power source to 100-110 V and let sit for approximately 30-45 minutes or until the dye marks are near the end of the gel.
- Turn of the power source, remove the gel, and use ultraviolet light to visualize the DNA bands.