Difference between revisions of "Team:UiOslo Norway/Experiments/Solubility Assay"
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Latest revision as of 12:19, 18 September 2015
Solubility Assay:
Back to ProtocolsCell 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:
