A | B | C | |
Reagent | For 1 gel (50 ml) | For 2 gels (100 ml) | |
Urea | 18 g | 36 g | |
40% polyacrylamide | 18.8 ml | 37.5 ml | |
10X TBE | 2.5 ml | 5 ml | |
NF water | Bring to 50 ml total volume | Bring to 100 ml total volume | |
Dissolve the above on stir plate. Assemble gel cassette(s), with comb in, and set in agar to seal bottom. Remove combs. When ready to cast gel, add: | |||
10% APS | 500 ul | 1 ml | |
Put on stir plate briefly to mix. | |||
TEMED | 50 ul | 100 ul | |
Put on stir plate briefly to mix, then promptly use 10 milliliter pipette to load. Carefully set combs, making sure there are no bubbles in the gel, especially where the wells will be formed. Get takes 5-15 minutes to polymerize. |
A | B | C | D | E | |
Reagent | grams | NF water (ml) | This makes stock concentration: | Notes | |
SDS | 1 | 10 | 10% | None | |
Bromophenol Blue | 0.25 | 10 | 2.5% | Pass through a 0.2 um syringe filter after dissolving | |
Xylene cyanol | 0.25 | 10 | 2.5% | Pass through a 0.2 um syringe filter after dissolving |
A | B | C | |
Recipe calls for: | Stock concentration: | Use this much of stock: | |
47.5% formamide | ultrapure (100%) | 4.75 ml | |
0.01% SDS | 10% | 10 ul | |
0.01% Bromophenol Blue | 2.5% | 40 ul | |
0.005% xylene cyanol | 2.5% | 20 ul | |
0.5 mM EDTA | 0.5 M | 10 ul |