Preparing a SDS-PAGE Gel: Difference between revisions
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" !...' |
added 6% gel |
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*Pour into cassette, overlay with the top layer of water saturated butanol | *Pour into cassette, overlay with the top layer of water saturated butanol | ||
{| border="1" | {| border="1" | ||
! Component || 7.5% || 10% || 12.5% | ! Component || 6% ||7.5% || 10% || 12.5% | ||
|- | |- | ||
| [[SDS-PAGE Separating Gel Buffer]] || 3 mL || | | [[SDS-PAGE Separating Gel Buffer]]|| 3 mL || 3 mL || 3 mL || 3 mL | ||
|- | |- | ||
| [[Acrylamide Solution]] || | | [[Acrylamide Solution]]|| 2.4 mL || 3 mL || 4 mL || 5 mL | ||
|- | |- | ||
| Water || 6 mL || 5 mL || 4mL | | Water || 6.6 mL|| 6 mL || 5 mL || 4mL | ||
|- | |- | ||
| APS (make fresh, 0.1g/mL in water) || 200 uL || 200 uL || 200 uL | | APS (make fresh, 0.1g/mL in water) || 200uL || 200 uL || 200 uL || 200 uL | ||
|- | |- | ||
| TEMED || 30 uL || 30 uL || 30uL | | TEMED || 30 uL || 30 uL || 30 uL || 30uL | ||
|} | |} | ||
*For a Phos-Tag Gel, add 24 uL 5 mM Phos-Tag and 24 uL 10 mM MnCl | |||
==Stacking Gel== | ==Stacking Gel== | ||