FCCT Biochemistry Lab:Agarose gel electrophoresis

From OpenWetWare
Jump to navigationJump to search
Igem-logo-150px.png iGEM Protocols Report.pngMain project page


Making Gel for gel electrophoresis

  • Make 100ml of gel at at time (e.g. 0.8%, 1%, 1.5%, or 2%)
  • Measure out the relevant percentage in agarose (e.g. 2g of agarose for 100ml of 2% gel).
  • Fill it up with 1xTAE buffer.
  • Microwave on low, constantly taking it out every so often to swirl and mix it. Keep microwaving until clear. Make sure it is completely clear
  • Cool to a temperature that is still hot but still can be held in your hand. If the gel gets too cold and starts to harden, start microwaving again until clear.
  • Add 3uL of ethidium bromide for every 100ml of gel after cooled to the correct tempearature. Mix it well by swirling
  • Pour it onto the molds quickly.
  • Wait at least 1/2hr until using it.

Gel electrophoresis

  • Select a relevant % agarose gel based on your own experience (general guide: all <1000: 2%, all >2000: 1% guide).
  • Load samples in wells.
  • Load 7uL of marker into a well.
  • Run in the 13x13cm box at 90V or 100V and 400mA for the desired amount of time.
  • View in the gel viewer machine.