Team:Tokyo Tech/Experiment/PHB2
From 2012.igem.org
Making purified P(3HB) sheets
We made P(3HB) sheets. To make the sheets, we cultured E.coli JM109 in erlenmeyer flasks at 37°C for 72h. Contact angle is an indicator to represent the strength of the water-repellent. The angle shows the physical properties, especially surface tension. When contact angle of sheets is larger than 90°, from Young equation, the sheets would have more strong water-repellent by increasing real surface area. Contact angle of P(3HB) sheets is about 100° from literature data. [Protocol]
Protocol
[Go to the P(3HB) production page "Application"]
1. Move dried cells into an airtight container.
2. Add chloroform, at rate of 2mg dried cells for 1 ml chloroform.
3. Incubate and stir the chloroform solution for more than 72 hrs at 20~25℃.
4. Filter the chloroform solution.
5. Concentrate by evaporation.
6. Dropwise the solution in methanol.
7. Filter the polymer in methanol, and dry the polymer.
8. Add a little chloroform to dissolve the polymer.
9. Poured the chloroform solution into a Petri dish.
10. Dry at room temperature.