Sheffield Molecular Genetics Facility
Making an agarose gel
1) Place masking tape at the ends of a 20cmx20cm plastic gel tray. Place four 27well combs in position. Place on a flat surface
2) Make up a 0.8% agarose gel (250ml 1xTBE, 1.75 g agarose) in a 500ml flask.
Heat in microwave (NB. take care as it becomes superheated. Stir every minute to
release the bubbles). Add 9ul ethidium bromide (1mg/ml) and cool under a tap
whilst rotating the flask
3) Pour into the prepared plastic gel tray
4) Leave to set for approx. 30 minutes
5) When set remove combs and place in electrophoresis tank containing 1XTBE
buffer
Preparing samples to load on an agarose gel
1) Pipette 14ul 1x Orange G loading buffer into wells into a 72-well Terrasaki plate
2) Add 1ul of the sample
3) To the last well add 0.5ug of Lambda DNA standard
4) Load all the sample on the gel
5) Electrophorese the gel (100Volts, DNA is –ve so goes toward the +ve electrode).