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Team:Columbia-Cooper-NYC/Gel Electrophoresis Setup
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Latest revision as of 03:40, 4 October 2012
Gel Electrophoresis Setup and Run Protocol
Note: The protocol is for creating 60ml of 1% agar gel used to run small DNA sequences of several hundred
To run gels for larger sequences (using the corresponding ladder), create a .7% agar gel
- Measure .6g of ultra pure agar
- Suspend agar in 60ml of TAE buffer
- Place cap on bottle containing the solution
- Set microwave for 120 seconds, let solution heat in microwave for first 45 seconds
- Take out bottle and shake bottle and place back in microwave
- Repeat step 5 every 10-15 seconds there after
- After microwaving let solution stand to cool in hood Note: Do not let gel solidify
- While cooling, rinse the gel mold with deionized water and make sure rubber gaskets are air tight and in place
- Place well molds in place
- Add .8µl Ethiolium bromide to TAE/agar mixture
- Pour the mixture into mold and let it stand until hardens
To run the gels:
- Gently remove wells from hardened gel
- For every 5µl of DNA add 1µl of 6x loading dye Note: Need to add loading dye to each sample of DNA)
- Add 7-8µl of 2 log ladder in first column
- Place desired DNA for inspection in all the other columns
- Run gel at steady 150V
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