Preparing a SDS-PAGE Gel

Revision as of 18:52, 3 August 2009 by Davebridges (Talk | contribs) (Created page with '==Separating Gel== *For 12 mL final volume (2 gels) *Combine, adding APS and TEMED last *Pour into cassette, overlay with the top layer of water saturated butanol {| border="1" !...')

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Revision as of 18:52, 3 August 2009 by Davebridges (Talk | contribs) (Created page with '==Separating Gel== *For 12 mL final volume (2 gels) *Combine, adding APS and TEMED last *Pour into cassette, overlay with the top layer of water saturated butanol {| border="1" !...')

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Separating Gel

  • For 12 mL final volume (2 gels)
  • Combine, adding APS and TEMED last
  • Pour into cassette, overlay with the top layer of water saturated butanol
Component 7.5% 10% 12.5%
SDS-PAGE Separating Gel Buffer 3 mL 4 mL 5 mL
Acrylamide Solution 3 mL 3 mL 3 mL
Water 6 mL 5 mL 4mL
APS (make fresh, 0.1g/mL in water) 200 uL 200 uL 200 uL
TEMED 30 uL 30 uL 30uL

Stacking Gel

  • Rinse polymerized separating gel with water
  • For 10 mL (2 gels)
  • Combine, adding APS and TEMED last
  • Pour over separating gel and insert comb
Component
SDS-PAGE Stacking Gel Buffer 2.5 mL
Acrylamide Solution 1.7 mL
Water 4 mL
APS (make fresh, 0.1g/mL in water) 200 uL 200 uL 200 uL
TEMED 30 uL 30 uL 30uL