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Preparing a SDS-PAGE Gel

939 bytes added, 18:52, 3 August 2009
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" !...'
==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"
! 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
{| border="1"
! 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
|}

[[Category: SDS-PAGE]]
[[Category: Electrophoresis]]