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Team:Tuebingen/Notebook/Protocols/SDS-PAGE
From 2014.igem.org
Protocols
SDS-PAGE
Separation Gel
| 4.5 mL | Acrylamid (40 %) |
| 3.75 mL | Tris-buffer pH 8.8 |
| 6.45 mL | H20 |
| 0.15 mL | SDS (10 %) |
| 6 µL | TEMED |
| 0.15 mL | APS (10 %) |
Stacking gel
| 0.75 mL | Acrylamid (40 %) |
| 0.76 mL | Tris-buffer pH 8.8 |
| 4.37 mL | H20 |
| 0.2 mL | SDS (10 %) |
| 6 µL | TEMED |
| 60 µL | APS (10 %) |
Procedure
- Mix all the reagents, adding APS and TEMED last (gives 2 gels).
- Fill the separation gel in the SDS page apparatus and overlay with isopropanol.
- After polymerization discard the isopropanol and fill the stacking gel on top and insert the comb.
- Preparation of the samples (add Laemmli-buffer).
- Load the gel and run at 200 V.
