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Large Scale Method: Sonication

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  1. Collect induced cells by centrifugation.
  2. Resuspend the cell pellet in 1/10 volume of 50 mM Tris-Cl pH 8, 2 mM EDTA.
  3. Add lysozyme to a concentration of 100 ug/mL (use a 10mg/mL stock prepared freshly in buffer from step 2). Add 1/10 volume 1% Triton X -100 Incubate 30 C 15 minutes.
  4. Sonicate sample in an ice bath using 2x10 sec. pulses at high output.
  5. Centrifuge at 12,000 g for 15 minute at 4C. The supernatant contains soluble proteins.