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Team:Paris Saclay/electro
From 2013.igem.org
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(Created page with "{{Team:Paris_Saclay/incl_debut_generique}} = '''Electro-Elution''' = 1. Cut the strip under the UV plate, cut the gel in little pieces 2. Fill the vat with TAE 0.5X 3. Close o...") |
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= '''Electro-Elution''' = | = '''Electro-Elution''' = | ||
| - | 1. Cut the strip under the UV plate, cut the gel in little pieces | + | 1. Cut the gel strip under the UV plate, then cut the gel in little pieces |
| - | 2. Fill the vat with TAE 0.5X | + | 2. Fill the vat with TAE 0.5X buffer |
| - | 3. Close one of the | + | 3. Close one of the wells of the saline bridge with a semi-permeable membrane, put the DNA in the large well |
4. Apply a 400V current after closing the vat, during 30min | 4. Apply a 400V current after closing the vat, during 30min | ||
| - | 5. Reverse the polarity during 20s, | + | 5. Reverse the polarity during 20s, pipet the solution from the small well and put it in a clean 1.5 ml tube |
| - | 6. Add 200µL of water and 600µL of phenol chloroform | + | 6. Add 200µL of water and 600µL of phenol/chloroform/isoamyl alcohol 25/24/1 |
| - | 7. Vortex during | + | 7. Vortex during 1 min |
| - | 8. | + | 8. Pipet the aqueous phase |
| - | 9. Precipitation with ethanol | + | 9. Precipitation the DNA with ethanol (see protocol) |
Revision as of 19:16, 3 October 2013
Electro-Elution
1. Cut the gel strip under the UV plate, then cut the gel in little pieces
2. Fill the vat with TAE 0.5X buffer
3. Close one of the wells of the saline bridge with a semi-permeable membrane, put the DNA in the large well
4. Apply a 400V current after closing the vat, during 30min
5. Reverse the polarity during 20s, pipet the solution from the small well and put it in a clean 1.5 ml tube
6. Add 200µL of water and 600µL of phenol/chloroform/isoamyl alcohol 25/24/1
7. Vortex during 1 min
8. Pipet the aqueous phase
9. Precipitation the DNA with ethanol (see protocol)
