Sample buffer preparation

Sample Buffer and Sample Preparation

Sample Buffer
Components
 Concentration
2x Buffer
 Amount
1 M Tris-HCl pH 6.8 0.126 M 0.625 ml
10 % (w/v) 4 % 2 ml
Glycerol 20 % 1 ml
0.1 % (w/v) Bromhenol blue 0.02 % 1 ml
2 M DTT 0.02 M 0 05 ml
Or: 2-Mercptoethanol 10 % 0.5 ml
Water, deion.   ad 5 ml

 

  • Since the reduction reagents oxidize in time, the buffer should always be freshly prepared. The samples are diluted (1:1) with an equal volume of 2x sample buffer and mixed well.

  • Heat sample for 5 minutes at 95 °C; heat fluorescence-labelled samples for 5 minutes at 65 °C.

  • Rinse wells with running buffer.

  • Load samples and start electrophoresis.

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