Sample buffer preparation
Sample Buffer and Sample Preparation
| Sample Buffer Components |
Concentration 2x Buffer |
Amount |
| 1 M Tris-HCl pH 6.8 | 0.126 M | 0.625 ml |
| 10 % (w/v) | 4 % | 2 ml |
| Glycerol | 20 % | 1 ml |
| 0.1 % (w/v) Bromhenol blue | 0.02 % | 1 ml |
| 2 M DTT | 0.02 M | 0 05 ml |
| Or: 2-Mercptoethanol | 10 % | 0.5 ml |
| Water, deion. | ad 5 ml |
- Since the reduction reagents oxidize in time, the buffer should always be freshly prepared. The samples are diluted (1:1) with an equal volume of 2x sample buffer and mixed well.
- Heat sample for 5 minutes at 95 °C; heat fluorescence-labelled samples for 5 minutes at 65 °C.
- Rinse wells with running buffer.
- Load samples and start electrophoresis.
