Preparation of bacteria-imprinted polymers

Caution This procedure should be carried out in a well-ventilated fume-cupboard with a UV shield in a laboratory equipped to Category 2 Microorganism Handling Standards. UV protective glasses, disposable gloves, and full length laboratory coat should be worn throughout. 

Obtain suspensions of microorganisms by overnight growth at 37 C of stock cultures in nutrient broth (S. enteritidis or coryneform broth L. monocytogenes.  

Centrifuge the stained cells (5 min, 3000 rpm), remove the supernatant, and re-suspend bacterial pellets in MOPS buffer (pH 7.8, 0.6 N, 10 ml) by vortex mixing (30 s). 

Stir a solution of MOPS buffer (pH 7.8, 0.6 N, 250 ml) in a reaction vessel (500 ml tail-form beaker) equipped with a magnetic bar at setting 5 over a IKA-MINI-MR stirrer plate whilst passing a steady stream of nitrogen via a Pasteur pipette for 10 min. 

Prepare a solution of adipoyl chloride (1.2 ml, 1.5 g, 8.2 mmol) in a mixed organic phase containing dibutyl ether (14.4 ml), 1,6-hexanediol diacrylate (14.4 ml, 14.5 g, 64 mmol), and AIBN (300 mg, 1.8 mmol), and add to the MOPS buffer. 

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