Team:UiOslo Norway/Experiments/Solubility Assay

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Solubility Assay:

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Cell lysis using glass beads

• Resuspend cell pellet in 600 µl PBS buffer (pH 7.4)

• Add glass beads and disrupt cells by vortexing

• Centrifuge at 16.000 x g for 5 minutes

• Transfer soluble fraction into a new tube and resuspend insoluble fraction in 600 ul PBS buffer (pH 7.4)

Cell lysis using sonication

• Resuspend the cell pellet in precooled imidazolebuffer.

• Sonicate 3x for 30 sec at a amplitude of 60. Incubate on ice for 1 minute between each sanitation step.

• Centrifuge the cell lysate at 13.000 rpm for 30 minutes at 4 °C.

• Keep the supernatant for further experiments

Imidazole buffer

50 mM Tris-HCl pH 8
100 mM NaCl
10 mM beta-Mercaptoethanol
10 mM Imidazole


iGEM UiOslo 2015 is sponsored by:

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