# Team:Tokyo Tech/Experiment/PHB2

(Difference between revisions)
 Revision as of 16:34, 26 October 2012 (view source)Takuo (Talk | contribs)← Older edit Revision as of 16:35, 26 October 2012 (view source)Takuo (Talk | contribs) (→D Make purified P(3HB) sheets)Newer edit → Line 12: Line 12: [[File:tokyotech PHB6.png|600px|thumb|center|Fig2-2-4-4-1, P(3HB) sheet]] [[File:tokyotech PHB6.png|600px|thumb|center|Fig2-2-4-4-1, P(3HB) sheet]] + ==Protocol== + [[http://2012.igem.org/Team:Tokyo_Tech/Projects/PHAs/index.htm#4-4_Make_purified_P.283HB.29_sheets Back to "4-4 Make purified P(3HB) sheets"]] + + + 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.

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# D Make 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]

Fig2-2-4-4-1, P(3HB) sheet

## Protocol

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.