Laemmli Sample Buffer (5X)

Reagents:

1. Weight 6.45 g SDS, add 16.125 ml 1M Tris, pH 7, add H2O to ~30 ml in a 50 ml Falcon tube.

2. Swirl the tube very slowly. You may warm it to 37C if necessary. It will take a while for SDS to be completely dissolved.

3. Add the solution into the DTT bottle and dissolve all DTT out. Add H2O to 32.25 ml. Swirl slowly to dissolve the DTT.

4. Transfer all solution to a 100 ml beaker and add 32.25 ml 100% glycerol.

5. Stir slowly until homogenous. Transfer several time between the beaker and a 50 ml Falcon tube to help.

6. Adjust pH to 6.8. The addition of DTT will affect pH of the solution.

7. Add 0.0645 g bromphenol blue to the beaker.

8. Aliquot and store at -20C with 1 ml each.


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