Equipment
- - Two glass plates
- - Gel caster
- - Comb
- - VWR Power Source 300V
Chemicals & consumables
- - SDS
- - Rotiphorese® (30%)
- - Tris HCl
- - Glycine
- - TEMED
- - APS
- - Aqua dest.
- - Isopropyle alcohol
Buffers & gels
- - Separation buffer (0.5 M Tris, 0.4% g SDS, pH = 8.8)
- - Stacking buffer (0.5 M Tris, 0.4% g SDS, pH = 6.6)
- - Running buffer (0.25 M Tris, 2 M Glycin, 1% SDS , pH = 8.3)
- - Separation gel 12.5% (5 ml Separation buffer, 6.25 ml Rotiphorese®, 3.75 ml Aqua dest., 30 µl TEMED, 30 µl APS (40%))
- - TEMED
- - APS
- - Aqua dest.
- - Isopropyle alcohol
Procedure
- prepare the separating gel and fill it into the chamber
- pour 1 mL isopropyle alcohol on the top of the gel to destroy air bubbles and prevent dehydration
- discard the isoprpyl alcohol and pour the prepared stacking gel
- stick in the comb
- store the gel in wet cloth (to prevent dehydration) at 4°C
"
