From 2007.igem.org

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• Applications: SDS PAGE

• Time to complete protocol:
  • Lab time: 15min, 5min.
  • Waiting time:30min, 5min.
• Approximate cost of materials: $

Contents 1 Method from primary and secondary reagents 1.1 Primary & secondary Reagents Required including controls 1.2 Method including controls 1.3 Equipement Required 1.4 References

Method from primary and secondary reagents

Primary & secondary Reagents Required including controls

For a 10% Gel:

• 4.2ml 30% acrylamide 1% bis
• 4.7ml 1M Tris pH 8.7
• 3.4ml water
• 125ul 10% SDS
• 62.5ul 10% Ammonium per sulfate
• 6.25ul TEMED

Stack:

• 0.83ml 30% acrylamide 1% bis
• 0.63ml 1M Tris pH 6.8
• 3.5ml water
• 50ul 10% SDS
• 25ul 10% Ammonium per sulfate
• 2.5ul TEMED

Method including controls

1. Set up Gel preparation apparutus by using 2 glass sheets, 2 spacers, a gel holder, and a rack.
2. In a conical flask, mix the Reagents excluding the ammonium per sulfate and the TEMED until ready to pour.
3. Pour the contents of the gel conical flask between the glass sheets using a pipette.
4. Wait for 30 minutes.
5. Mix the ammonium per sulfate and TEMED into the stack flask and pipette it between the glass sheets, on top of the gel.
6. Insert comb between the sheets.
7. Wait till the stack settles (5min) before loading the gel. Alternatively, gel can be stored up to a week in the fridge.

Equipement Required

• Measuring cylinders
• Pipette and tips
• Gel preparation apparatus and comb

References