Oneill Lab:Gels
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Michael J. O'Neill Lab University of Connecticut Department of Molecular and Cell Biology
Agarose gels
- Electrophoresis grade agarose gels are made with TBE
- NuSieve gels for DNA purification are made with TAE
Formaldehyde gel
For making a 1% 100 mL
- Combine
- 5mL 20x Running Buffer
- 77mL deionized water
- 1g agarose
- Boil in microwave until agarose dissolves
- Add 18mL 37% formaldehyde
- Swirl flask while pouring
- Add formaldehyde in fume hood
- Pour gel in fume hood
Polyacrylamide gels
10% | 7.5% | 5% | |
---|---|---|---|
Acrylamide | 8.3 ml | 6.25 ml | 4 ml |
1x TBE | 16.4 ml | 18.5 ml | 20.8 |
10% APS | 250 μL | 250 μL | 250 μL |
TEMED | 25 μL | 25 μL | 25 μL |
5 ml stacking gel (5%) 20 ml resolving gel |
5% | 8% | 9% | 10% | 11% | 12% |
---|---|---|---|---|---|---|
Range of separation (kD) | 40-100 | 30-90 | 20-80 | 16-70 | 12-60 | |
H2O (ml) | 9.3 | 8.6 | 7.9 | 7.2 | 6.5 | |
29:1 Acrylamide:bis-acrylamide (Protogel) |