Team:Edinburgh/Notebook/Blue light producer

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   <li><a href="http://2010.igem.org/Team:Edinburgh/BioBricks#Genomic">submitted parts</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/BioBricks#Genomic">submitted parts</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Results#Genomic">results</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Results#Genomic">results</a></li>
-
   <li><a href="http://2010.igem.org/Team:Edinburgh/Project/Future">future work</a></li>
+
   <li><a href="http://2010.igem.org/Team:Edinburgh/Project/Future">the future</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Project/References">references</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Project/References">references</a></li>
   </ul>
   </ul>
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  <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial" class="dir">bacterial BRIDGEs</a>
  <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial" class="dir">bacterial BRIDGEs</a>
   <ul>
   <ul>
-
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Core_repressilator">the repressilator</a></li>
+
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Core_repressilator">the project</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_producer">red light</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_producer">red light</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_sensor">red sensor</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_sensor">red sensor</a></li>
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   <li><a href="http://2010.igem.org/Team:Edinburgh/BioBricks#Bacterial">submitted parts</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/BioBricks#Bacterial">submitted parts</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Results#Bacterial">results</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Results#Bacterial">results</a></li>
-
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Future">future work</a></li>
+
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Future">the future</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/References">references</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Bacterial/References">references</a></li>
   </ul>
   </ul>
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   <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/Bacterial">the bacterial model</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/Bacterial">the bacterial model</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/Signalling">the signalling model</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/Signalling">the signalling model</a></li>
 +
  <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/Tools">tools</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Results#Modelling">results</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Results#Modelling">results</a></li>
-
   <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/Future">future work</a></li>
+
   <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/Future">the future</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/References">references</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Modelling/References">references</a></li>
   </ul>
   </ul>
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  <li><a href="http://2010.igem.org/Team:Edinburgh/Human" class="dir">human BRIDGEs</a>
  <li><a href="http://2010.igem.org/Team:Edinburgh/Human" class="dir">human BRIDGEs</a>
   <ul>
   <ul>
-
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Aspects">human aspects</a></li>
+
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Communication">communication of science</a></li>
 +
  <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Branding">iGEM survey</a></li>
 +
  <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Conversations">conversations</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Epic">the epic</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Epic">the epic</a></li>
-
   <li><a href="http://2010.igem.org/Team:Edinburgh/Results#Human">results</a></li>
+
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/FutureApps">future applications</a></li>
-
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Future">future work</a></li>
+
   <li><a href="http://2010.igem.org/Team:Edinburgh/Results#Human">further thoughts</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/References">references</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/References">references</a></li>
   </ul>
   </ul>
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   <ul>
   <ul>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Collaboration">collaboration</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Collaboration">collaboration</a></li>
 +
  <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Attribution">attribution</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/BRIDGE">BRIDGE</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/BRIDGE">BRIDGE</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Red_light_producer">red light</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Red_light_producer">red light</a></li>
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   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Blue_light_producer">blue light</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Blue_light_producer">blue light</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Blue_light_sensor">blue sensor</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Blue_light_sensor">blue sensor</a></li>
 +
  <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Green_light_producer">green light</a></li>
 +
  <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Green_light_sensor">green sensor</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Modelling">modelling</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Modelling">modelling</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Safety">safety</a></li>
   <li><a href="http://2010.igem.org/Team:Edinburgh/Notebook/Safety">safety</a></li>
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<br>
<br>
<br>
<br>
 +
 +
<div id="windowbox" style="border: .2em solid #660000; padding: 5px; position:fixed; top:50%; right:30px; width:8%;">
 +
<span style="color:ivory;">These <b>notes</b> correspond to the part of the <b>project</b> detailed <a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Blue_light_producer">here</a>.</span>
 +
 +
</div>
</html>
</html>
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== Blue Light Producer ==
== Blue Light Producer ==
<br>
<br>
 +
'''5/7/2010'''
 +
*LuxAB & LuxCDE operon agarose gel
 +
* Lane1: About2.1.kbp
 +
* Lane 2: The highest band- very faint- about 3.5kbp
 +
[[Image:150.JPG]]
 +
'''13/7/2010'''
'''13/7/2010'''
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*The transformations of Edinbrick are fine but all the luxABs failed (grew blue, not white on Xgal and did not glow, as of 27/7)/
*The transformations of Edinbrick are fine but all the luxABs failed (grew blue, not white on Xgal and did not glow, as of 27/7)/
*Still have some digested LuxAB-ediI so re-did ligation and transformation.  
*Still have some digested LuxAB-ediI so re-did ligation and transformation.  
-
'''Gel Pic- minipreps'''
+
[[Image:154.JPG]]
{| border="1"
{| border="1"
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| LuxAB  
| LuxAB  
|}
|}
 +
[[Image:155.JPG]]
 +
{| border="1"
 +
| Lane 1
 +
| Lane 2 +3
 +
| Lanes 3-6
 +
|-
 +
| Ladder
 +
| EdiI
 +
| LuxAB
 +
|}
 +
 +
'''29/7/2010'''
 +
 +
[[Image:156.jpg]]
 +
 +
* The ligation had the same gel characteristics as previous luxAb-ediI ligations, e.x. it isn't visivle- not hopeful of this transformation working
 +
 +
'''4/8/2010'''
 +
* made minipreps of blue (negative colonies)- double digest and ran in comparison with pure edinbrickI
 +
* LacZ band dissapears but colonies still grow blue (WTF)
 +
* Has bands equaling to luxAB and EdiI but there was no glowing
 +
 +
[[Image:162.jpg]]
 +
 +
'''12/8/2010''' 
 +
update on LumP:
 +
* still getting pink/red colonies from transformants
 +
* single digests give a band of about 3kb
 +
* double digests give a band of about 2.5 kb -highly confusing
 +
* and also, no sign on RFP despite red/pink growth.
 +
* Vector wa ssequenced and LumP is definately there.
 +
 +
[[Image:160.jpg]]
 +
 +
Lanes 2-5: single digests of lumP
 +
Lane 6: double digest of lumP
 +
 +
'''26/8/2010'''
 +
 +
* Marker (Lane 1), minipreps 6-10 luxABlumP (lanes 2-6)
 +
[[Image:129.JPG]]
 +
* single digests of LuxABlumP
 +
[[Image:131.JPG]]
 +
 +
'''7/9/2010'''
 +
*minipreps of luxCDE: samples 1&4 usable
 +
 +
{| border="1"
 +
| Lane 1
 +
| Lane 2-6
 +
| Lanes 7
 +
|-
 +
| Ladder
 +
| luxCDE00:1-5 minipreps
 +
| Ladder
 +
|}
 +
[[Image:140.JPG]]

Latest revision as of 02:12, 28 October 2010





These notes correspond to the part of the project detailed here.

Blue Light Producer


5/7/2010

  • LuxAB & LuxCDE operon agarose gel
  • Lane1: About2.1.kbp
  • Lane 2: The highest band- very faint- about 3.5kbp

150.JPG

13/7/2010

Empty edinbrick vector for control.

  • LuxAB + edinbrick ligation.

28/7/2010

an update on LuxAB:

  • The transformations of Edinbrick are fine but all the luxABs failed (grew blue, not white on Xgal and did not glow, as of 27/7)/
  • Still have some digested LuxAB-ediI so re-did ligation and transformation.

154.JPG

Lane 1&15 Lane 2 Others
Ladder EdiI LuxAB

155.JPG

Lane 1 Lane 2 +3 Lanes 3-6
Ladder EdiI LuxAB

29/7/2010

156.jpg

  • The ligation had the same gel characteristics as previous luxAb-ediI ligations, e.x. it isn't visivle- not hopeful of this transformation working

4/8/2010

  • made minipreps of blue (negative colonies)- double digest and ran in comparison with pure edinbrickI
  • LacZ band dissapears but colonies still grow blue (WTF)
  • Has bands equaling to luxAB and EdiI but there was no glowing

162.jpg

12/8/2010 update on LumP:

  • still getting pink/red colonies from transformants
  • single digests give a band of about 3kb
  • double digests give a band of about 2.5 kb -highly confusing
  • and also, no sign on RFP despite red/pink growth.
  • Vector wa ssequenced and LumP is definately there.

160.jpg

Lanes 2-5: single digests of lumP Lane 6: double digest of lumP

26/8/2010

  • Marker (Lane 1), minipreps 6-10 luxABlumP (lanes 2-6)

129.JPG

  • single digests of LuxABlumP

131.JPG

7/9/2010

  • minipreps of luxCDE: samples 1&4 usable
Lane 1 Lane 2-6 Lanes 7
Ladder luxCDE00:1-5 minipreps Ladder

140.JPG