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Team:Northwestern/preparegel
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Latest revision as of 00:39, 28 September 2013
Preparing Gel
Materials:
- agarose
- 0.5X TBE
- ethidium bromide
- For most procedures, a 1% agarose gel is sufficient. If you need to separate smaller pieces of DNA fragments (<1 kb) you may wish to use a 1.5% or 2% gel. If you wish to separate larger fragments, you may wish to use a 0.8% gel.
- A 65mL solution for a 1.5% gel requires 0.975g of agarose. Mix this with 65mL of 0.5X TBE buffer in a 125mL erlenmeyer flask.
- Microwave in order to dissolve the agarose in the buffer. If the buffer is allowed to boil, it with froth out of the flask, so stop the microwave before this happens. Take the flask out after each stop and swirl it to mix. Do this until the agarose is completely dissolved. CAUTION: the agarose turns transparent, and so may appear to be dissolved when it is not. Bring it up to the light to make sure it is homogenous.
- Add 1uL of ethidium bromide for every 10mL of gel solution. WARNING: Ethidium bromide is mutagenic, so take proper precautions. Wear gloves and goggles. Dispose of the tip in the ethidium bromide trash (yellow trash bag).
- Allow to cool until you can touch the sides for 4-5 seconds without glo
