Kay Finn
last updated: June 12, 2002
7X Native Gel Upper (Stacking)
Buffer:
5.7 g Tris base, pH to 6.7 with H3PO4
water to 100 ml total vol
4X Native gel Lower (Separating)
Buffer:
18.2 g Tris Base, pH to 8.9 with HCl
water to 100 ml
50X Running Buffer:
7.5 g Tris base
36 g Glycine
water to 250 ml
3X Sample Buffer:
3 ml glycerol
0.6 ml 50X Running Buffer
6.4 ml H2O
bromophenol blue
Sample gel recipes:
Separating Layer (Lower; 7.5%)
6 ml H2O
3 ml Lower buffer
3 ml Acrylamide/Bis (30/0.8)
7 ul TEMED
84 ul 10% APS
Stacking Layer (Upper;
4.3%)
5 ml H2O
1 ml upper buffer
1 ml Acrylamide/Bis (30/0.8)
8 ul TEMED
40 ul 10% APS
Using Biorad Criterion
cassettes (cat # 3459902):
10 ml vol for separating layer and level with H2O, set for 1 hr
Pour off water and dry with filter paper,
~4 ml for upper layer; insert comb and wait for 1hr
Wrap with saran if not using same day, and store in cold room.