Team:Edinburgh/Notebook/Blue light producer

From 2010.igem.org

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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/Tools">tools</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/Communication">communication of science</a></li>
-
   <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Terminology">terminology research</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/Wiki">wiki</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/Conversations">conversations</a></li>
-
  <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Identity">identity</a></li>
 
-
  <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Collaboration">collaboration</a></li>
 
-
  <li><a href="http://2010.igem.org/Team:Edinburgh/Human/Branding">branding research</a></li>
 
-
  <li><a href="http://2010.igem.org/Team:Edinburgh/Human/SciFi">science fiction writing</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/SelfReflection">self-reflection</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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<div id="windowbox" style="border: .2em solid #660000; padding: 5px; position:fixed; top:50%; right:30px; width:8%;">
<div id="windowbox" style="border: .2em solid #660000; padding: 5px; position:fixed; top:50%; right:30px; width:8%;">
-
<span style="color:ivory;">Throughout this wiki there are words in <b>bold</b> that indicate a relevance to <b>human aspects</b>. It will become obvious that <b>human aspects</b> are a part of almost everything in <b>iGEM</b>.</span>
+
<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>
</div>
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| LuxAB  
| LuxAB  
|}
|}
-
[[Image:154.JPG]]
+
[[Image:155.JPG]]
{| border="1"
{| border="1"
| Lane 1
| Lane 1
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| LuxAB  
| 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
* The ligation had the same gel characteristics as previous luxAb-ediI ligations, e.x. it isn't visivle- not hopeful of this transformation working
-
'''29/7/2010'''
+
'''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'''   
'''12/8/2010'''   
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* Vector wa ssequenced and LumP is definately there.  
* Vector wa ssequenced and LumP is definately there.  
-
'''gel pohot no x and y''' + description.
+
[[Image:160.jpg]]
 +
 
 +
Lanes 2-5: single digests of lumP
 +
Lane 6: double digest of lumP
'''26/8/2010'''
'''26/8/2010'''

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