C.B.S. Scientific MGV-402 - Manual (Page 15)

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4.2.1 Recommended Buffers and Reagents
Pre-mix acrylamide stock solutions are the method of choice. Use according to
manufacturer’s instructions. All C.B.S. Scientific Mini-Vertical units will accommodate
commercially available pre-cast gels.
Typical ‘scratch’ recipe for a 4% acrylamide mini-gel:
2.5 mls 40% acrylamide; 2% bis stock
2.6 mls 1.0M Tris
4.9 mls dH20
100 ul 10% APS
5-10 ul TEMED
1. Make up 0.5X buffer
2. After gel apparatus is set-up and ready for the gel to be poured, add 100ul
fresh 10% APS solution.
3. Add 5 to 10ul TEMED and using a pipette, quickly “pour” the gel.
4. Allow the gel to polymerize at least 20 to 30 minutes.
5. Remove the comb after polymerization and wash out wells with 0.5X TBE
(acrylamide will seep into the wells).
6. Fill upper and lower chambers with 0.5X TBE
7. Pre-electrophorese gel, if needed.
8. Load wells with samples.
9. Run at 75 VDC to 125VDC constant approximately 2 hours.
10. Monitor migration with dye markers.
BUFFERS:
Agarose Page/Agarose Slab
TBE (1X solution): TAE (1X):
0.089M Tris base
0.04M Tris-acetate
0.89 M Boric acid 0.001M EDTA
0.002M EDTA pH 8.0
pH 8.3
Protein Denaturing Denaturing/Non-Denaturing/
TG-SDS(1X):
Nylon Blotting
0.025M Tris base TT (Tris-Taurine( (1X): TT-SDS(1X):
0.192M Glycine 0.1M Tris base 0.1M Tris base
0.1%(w/v) SDS 0.1M Tricine 0.1M Tricine
pH 8.3 0.1%(w/v) SDS