TITLE: RAFT Based Synthesis of In-house Polymers
AUTHORS: Caroline Brown, Snehasish Ghosh, Kallol Gupta

[DESCRIPTION]
The protocol gives details for synthesizing the ChloroSMA series of membrane active polymers.

[STEPS]
SECTION: Chemicals and reagents
1. All reagents were purchased from Sigma-Aldrich and used without further purification unless specified 
otherwise. 
SECTION: Synthesis of ChloroSMA20
2. To a 50 mL add 1.52 millimolar (mM) ,1.52 millimolar (mM) , and 0.076 millimolar (mM) .

SECTION: Synthesis of ChloroSMA 40
8. To a 50 mL add 3.1 millimolar (mM) ,3.1 millimolar (mM) , and 0.076 millimolar (mM). 


SECTION: Synthesis of ChloroSMA20
3. Dissolve in dimethylformamide.
SECTION: Synthesis of ChloroSMA20
4. Add 0.004 millimolar (mM) .
SECTION: Synthesis of ChloroSMA20
5. Cap the flask with a rubber stopper and bubble with nitrogen for 15 min  minutes.

SECTION: Synthesis of ChloroSMA20
6. Heat to 90 °C  for 960 min hours.

SECTION: Synthesis of ChloroSMA20
7. Precipitate in isopropanol or water, filter, and dry in vacuo.
SECTION: Synthesis of ChloroSMA 40
9. Dissolve in dimethylformamide.
SECTION: Synthesis of ChloroSMA 40
10. Add 0.004 millimolar (mM) .
SECTION: Synthesis of ChloroSMA 40
11. Cap the flask with a rubber stopper and bubble with nitrogen for 15 min  minutes.
SECTION: Synthesis of ChloroSMA 40
12. Heat to 90 °C  for 960 min hours.
SECTION: Synthesis of ChloroSMA 40
13. Precipitate in isopropanol or water, filter, and dry in vacuo.
SECTION: Synthesis of ChloroSMA 60
14. To a 50 mL add 4.56 millimolar (mM) ,4.56 millimolar (mM) , and 0.076 millimolar (mM) .

SECTION: Synthesis of ChloroSMA 60
15. Dissolve in dimethylformamide.
SECTION: Synthesis of ChloroSMA 60
16. Add 0.004 millimolar (mM) .
SECTION: Synthesis of ChloroSMA 60
17. Cap the flask with a rubber stopper and bubble with nitrogen for 15 min minutes.

SECTION: Synthesis of ChloroSMA 60
18. Heat to 90 °C  for 960 min hours.
SECTION: Synthesis of ChloroSMA 60
19. Precipitate in isopropanol or water, filter, and dry in vacuo.
SECTION: Synthesis of ChloroSMA 80
20. To a 50 mL add 6.1 millimolar (mM) ,6.1 millimolar (mM) , and 0.076 millimolar (mM) .
SECTION: Synthesis of ChloroSMA 80
21. Dissolve in dimethylformamide.
SECTION: Synthesis of ChloroSMA 80
22. Add 0.004 millimolar (mM) .
SECTION: Synthesis of ChloroSMA 80
23. Cap the flask with a rubber stopper and bubble with nitrogen for 15 min minutes.

SECTION: Synthesis of ChloroSMA 80
24. Heat to 90 °C  for 960 min hours.
SECTION: Synthesis of ChloroSMA 80
25. Precipitate in isopropanol or water, filter, and dry in vacuo.
SECTION: End-group removal of poly(4 chlorostyrene-alt-maleic anhydride
26. Dissolve precipitated polymer in dimethylformamide and add 9.9 millimolar (mM)  to a round bottom flask.

SECTION: End-group removal of poly(4 chlorostyrene-alt-maleic anhydride
27. Seal with a rubber stopper and bubble with nitrogen for 15 min  minutes.

SECTION: End-group removal of poly(4 chlorostyrene-alt-maleic anhydride
28. Leave the escape needle in the flask and heat to 90 °C for 360 min  hours.

SECTION: End-group removal of poly(4 chlorostyrene-alt-maleic anhydride
29. Precipitate polymer with isopropanol or water and dry in vacuo.
SECTION: Hydrolysis to produce poly(4 chlorostyrene-alt-maleic acid)
30. In a round bottom flask dissolve 800 mg  in 2 g and 20 mL.

SECTION: Hydrolysis to produce poly(4 chlorostyrene-alt-maleic acid)
31. Reflux for 240 min hours. The mixture should become clear.

SECTION: Hydrolysis to produce poly(4 chlorostyrene-alt-maleic acid)
32. Dialyze the hydrolyzed polymer using a 3.5KDa mwco membrane for 2160 min hours. 

SECTION: Hydrolysis to produce poly(4 chlorostyrene-alt-maleic acid)
33. Add water to precipitate the polymer.
SECTION: Hydrolysis to produce poly(4 chlorostyrene-alt-maleic acid)
34. Wash polymer using 0.1N HCl, 3 times, and dry the polymer under nitrogen.