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Sorbitol/Betaine Method

- The use of osmotic stress to facilitate the uptake of the 'compatible solute', glycyl betaine. 

- reference --Blackwell, JR. and R. Horgan (1991) FEBS Letters vol 295 p10-12. 

Grow in media containing 2.5 mM Betaine and try various amounts of Sorbitol (330 mM, 660 mM, 1 M for instance) and check for inclusion body production. Filter sterilize Betaine and Sorbitol solutions and add to appropriately concentrated autoclaved LB (reason: not sure if Sorbitol or Betaine can be autoclaved).

3.3 M Sorbitol

60.6 g in 100 mL water

0.5 M Betaine

7.08g in 100 mL