Haynes Lab:Notebook/Engineering PC-TFs/2012/11/08

• Gel Electrophoresis

Follow steps for gel electrophoresis.

• Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray.
• Create 1% gel by putting .6 grams of agarose into flask.
• Microwave agarose solution for 30 seconds
• Using hot gloves, grab flask out of microwave and swirl flask vigorously. Place back in microwave for 30 more seconds.
• When flask is taken out of microwave, make sure that the agarose is completed dissolved.
• Set up wells on gel electrophoresis apparatus. The mold for the first digest is B-10. Use the thin "teeth" and slip in the notches.
• Add EtBr (ethidium bromide) to the agarose gel. There should be 1 microliter of EtBr per 10ml of agarose gel. Therefore, 6 µL.
• Pour gel into tray. Wait twenty minutes for gel to settle.
• Wash the agarose gel flask.
• Pipette 15 μL of DNA ladder into first well. Pipette 5μL of PCR reactions into subsequent wells.
• Turn on electrophoresis and let run for thirty minutes.

The assembly did not work. No bands were seen when placed under UV light.