Berglund:Agarose

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Contents

1% Agarose Gel

Materials

  • 1g of molecular biology grade (Shelton scientific-IB70042) agarose
  • 100ml of 1xTAE buffer
    • 10xTAE
      • 193.6g Tris
      • 45.68ml Glacial Acetic Acid
      • 80ml 0.5M EDTA
      • 3874ml water
  • 10mg/ml Ethidium bromide

Methods

  1. Heat carefully, swirling often in a microwave (I recommend a bottle that holds 500ml and is either a flask or a screw-top). Can also be put in a 65°C water bath and kept there to be used when desired.
  2. After heating add:
    • 7µl of 10mg/ml Ethidium bromide
  3. Swirl, allow to cool a little or the Plexiglas frame of your gel apparatus will warp and (definitely) crack overtime.
  4. Place comb into slots on gel apparatus.
  5. Pour a thin layer (approximately 6mm) (30ml) for a 12 well gel.
  6. Cool for 10-20 minutes (until gel is completely hardened).
  7. Pour fresh 1x TAE into the apparatus until just over the gel surface (the more buffer above your gel the more current that does not go through the gel and the longer it takes for your gel to run).
  8. Load the samples and appropriate ladder, run at 100-150V.
  9. View using a UV light box and take a picture.


3% Agarose Gel

Same as above but with 3g of agarose to 100ml of 1xTAE. You need to be very careful about boiling this in the microwave, it explodes out the top and makes a mess, so stopping and starting the microwave with mixing in between stops is necessary. Also the little bubbles that never go away can be a problem in your pictures, but if you let the molten agarose harden and then reheat it behaves beautifully.

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