User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/09/25

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Lysis of transformed E. coli

  • To retrieve the adenosine deaminase (ADA) from E.coli, the cell must be lysed. By lysing the cell, the cytoplasm will no longer isolate the macromolecules, such as the ADA for this instance, from the extracellular environment.
  • Samples 1 and sample 2 which were placed below -80°C on Sept. 19, 2012 is taken out from the freezer to thaw to room temperature.
  • There are different methods by which the cell can be lysed. These methods include heat shock, blender polytron, homogenization etc. For this procedure, sonication will be used to lyse the cell. By exposing the cells to high frequency sound waves of a sonicator, the cell membrane are sheared exposing the intracellular contents.
  • The sonicator used is the Misonix XL-2000 series Ultrasonic Liquid Processors.


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