User:Tamra L. Fisher/Notebook/Experimental Biological Chem/2011/09/20

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Objective

Extract protein from ER2566 E.coli with the pMAL-pIII plasmid. Begin the purification of the maltose binding protein (MBP).

Description

Extraction

  1. Defrost cells that were stored at -20°C.
  2. Sonicate the cells for 15 seconds at a time with approximately one minute on ice between sonications. In total the cells were sonicated for 90 seconds.
  3. Centrifuge the cells at 9000g for 30 minutes at 4°C.

Purification

  1. The column was poured. Amylose resin was used. The column volume was 15mL.
  2. 120mL of the column buffer was run over the column at 2mL/min.