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Team:HokkaidoU Japan/Notebook/plastic protocols
From 2012.igem.org
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Revision as of 10:30, 6 August 2012
Polymer producing media
polymer producing media (LB:2%Glc+ 10mM pantothenic acid Ca + Amp100mg/l) 20ml
| 2×LB | 10ml |
| ●50% Glucose | 800ul |
| ●1M pantothenic acid Ca | 200ul |
| Amp(100mg/ml) | 20ul |
| ROwater | 8.98ml |
Put 1.5ml each into the test tube.
●50%gulcose
| ROwater | 7ml |
| Glucose | 10g....Heat and stir until it melts. |
| ROwater | up to 20ml |
Filter sterilize.
●1M pantothenic acid Ca
| ROwater | 7ml |
| pantothenic acid Ca | 4.77g....Heat and stir until it melts. |
| ROwater | up to 10ml |
Filter sterilize.
Culture and harvest
Polymer extraction and purification
<p>
- Move dried up bacteria into test tube.
- Break up them to separate and add 10ml chloroform.
- Incubate for 48hour at 60C.
- Make it filtered through PTFE and move it into another test tube.
- Volatilize organic solvent by exposing air and separate polymer.
- Add 5ml hexane and voltex for a minute. After centrifuging (1,500rpm, 10min), remove the clear layer.
- Volatilize chloroform by exposing air again.
- Add 5ml MtOH, voltex for a minute, centrifuge (1,500rpm, 10min), and remove the clear layer.
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