Team:HokkaidoU Japan/Notebook/plastic protocols

From 2012.igem.org

(Difference between revisions)
(→Polymer extraction and purification)
Line 3: Line 3:
<div id="hokkaidou-column-main">
<div id="hokkaidou-column-main">
<!-- DO NOT EDIT OVER THIS LINE @iTakeshi -->
<!-- DO NOT EDIT OVER THIS LINE @iTakeshi -->
 +
 +
===Polymer producing media===
 +
<div>
 +
<p>
 +
polymer producing media (LB:2%Glc+ 10mM pantothenic acid Ca + Amp100mg/l) 20ml
 +
{|class="hokkaidou-table-digestion"
 +
|-
 +
|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.
 +
 +
polymer producing media (LB:2%Glc+ 10mM pantothenic acid Ca + Amp100mg/l) 20ml
 +
{|class="hokkaidou-table-digestion"
 +
|-
 +
|2×LB
 +
|10ml
 +
|-
 +
|50% Glucose
 +
|800ul
 +
|-
 +
|1M pantothenic acid Ca
 +
|200ul
 +
|-
 +
|Amp(100mg/ml)
 +
|20ul
 +
|-
 +
|ROwater
 +
|8.98ml
 +
|-
 +
|}
 +
 +
</p>
 +
===Polymer extraction and purification===
===Polymer extraction and purification===

Revision as of 09:19, 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.

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


Polymer extraction and purification

  1. Move dried up bacteria into test tube.
  2. Break up them to separate and add 10ml chloroform.
  3. Incubate for 48hour at 60C.
  4. Make it filtered through PTFE and move it into another test tube.
  5. Volatilize organic solvent by exposing air and separate polymer.
  6. Add 5ml hexane and voltex for a minute. After centrifuging (1,500rpm, 10min), remove the clear layer.
  7. Volatilize chloroform by exposing air again.
  8. Add 5ml MtOH, voltex for a minute, centrifuge (1,500rpm, 10min), and remove the clear layer.



Retrieved from "http://2012.igem.org/Team:HokkaidoU_Japan/Notebook/plastic_protocols"