Team:Edinburgh/Notebook/Blue light producer

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<div id="nav">
<div id="nav">
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<ul class="dropdown dropdown-horizontal">
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<ul class="dropdown dropdown-horizontal" style="background: none;">
  <li><a href="http://2010.igem.org/Team:Edinburgh" class="dir">home</a>
  <li><a href="http://2010.igem.org/Team:Edinburgh" class="dir">home</a>
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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>
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   <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>
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   <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>
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   <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>
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<div id="windowbox" style="border: .2em solid #660000; padding: 5px; position:fixed; top:50%; right:30px; width:8%;">
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<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>
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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Mauris auctor dolor quis diam fringilla non malesuada sapien volutpat. Proin purus arcu, tristique ut ultricies eget, elementum ac odio. Nam gravida blandit nisl, in vestibulum nunc malesuada id. Cras sit amet mi a massa aliquet auctor. Morbi lorem magna, ultrices vitae iaculis id, malesuada vitae nibh. Fusce elementum euismod vehicula. Maecenas mattis bibendum elit, sed semper diam scelerisque eget. Vivamus enim arcu, consequat non volutpat nec, hendrerit id quam. Quisque dapibus, augue non scelerisque tincidunt, ipsum magna posuere nunc, consectetur blandit elit odio non orci. Morbi congue molestie ipsum non ultricies. Pellentesque sed nisl nunc, id facilisis magna. Sed sit amet est ut magna tempor eleifend. Duis pulvinar lobortis rutrum.
+
== Blue Light Producer ==
 +
<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'''
 +
 
 +
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.  
 +
[[Image:154.JPG]]
 +
 
 +
{| border="1"
 +
| Lane 1&15
 +
| Lane 2
 +
| Others
 +
|-
 +
| Ladder
 +
| EdiI
 +
| 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
-
Etiam pretium mattis enim, ac pellentesque quam congue at. Ut interdum, velit sed aliquam dapibus, lacus turpis gravida nibh, sed iaculis sem ligula ut mi. Maecenas facilisis volutpat elit, sit amet pharetra dolor porta ac. Proin posuere, ante ac pretium luctus, ante nunc luctus orci, at mattis mi sem sit amet nisi. Sed venenatis auctor mi, ac porttitor quam ultricies quis. Suspendisse at orci elit. Praesent egestas metus at sem lacinia eu hendrerit est convallis. Praesent cursus mauris purus, id blandit nibh. Nulla consectetur, nunc in ornare mattis, ligula massa gravida magna, a pellentesque metus lacus id nunc. Donec eget ipsum eu turpis placerat semper. Donec ligula turpis, lobortis id sodales eget, fermentum vel turpis. Aenean at convallis eros. Cras cursus ligula id arcu venenatis accumsan sit amet volutpat nulla. Donec id ante id nisi porta volutpat sit amet quis quam. Vivamus tincidunt ligula id ante mollis non auctor sem porta. Etiam eleifend laoreet nunc vel laoreet. Cras quis nunc ac mi pulvinar imperdiet. Nam nec arcu non tortor tincidunt commodo. Cum sociis natoque penatibus et magnis dis parturient montes, nascetur ridiculus mus.
+
'''26/8/2010'''
-
In hac habitasse platea dictumst. Praesent at dolor arcu, non cursus justo. Fusce non nibh felis. Nulla pharetra eleifend egestas. Sed semper ligula ac mi tincidunt mollis. Ut elementum justo sed urna luctus sed consectetur mauris euismod. Suspendisse potenti. Phasellus quam sem, ultrices et vulputate eu, egestas vitae nisi. Morbi non tellus sapien, faucibus sagittis diam. In hac habitasse platea dictumst. Aliquam tempor ipsum at sapien lobortis viverra. Sed volutpat nunc in justo vehicula egestas. Vestibulum ante ipsum primis in faucibus orci luctus et ultrices posuere cubilia Curae; Nullam sed nisi velit. Praesent at quam nec metus imperdiet tincidunt vitae eget mi. Phasellus laoreet tincidunt tempus. Fusce nisl enim, hendrerit quis consequat quis, vestibulum pharetra lectus. Morbi cursus gravida sem, interdum sollicitudin mi aliquam id. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Morbi eleifend hendrerit leo, nec aliquet elit adipiscing vitae.
+
* Marker (Lane 1), minipreps 6-10 luxABlumP (lanes 2-6)
 +
[[Image:129.JPG]]
 +
* single digests of LuxABlumP
 +
[[Image:131.JPG]]
-
Ut commodo vehicula erat eget laoreet. Duis elit quam, malesuada id semper sit amet, bibendum pellentesque risus. Nam non lacus nunc. Fusce mi arcu, fermentum eu tempus in, ornare sit amet sapien. Donec fringilla sem vel nunc hendrerit sit amet mollis urna pharetra. Duis eu arcu nec eros tempus interdum. Mauris tempus ultrices sapien ut malesuada. Fusce erat turpis, bibendum nec rutrum ac, dapibus id erat. In pretium malesuada libero, eu commodo purus sagittis in. Mauris est nibh, pulvinar in vulputate sed, porttitor sit amet nisl. Sed mattis fermentum erat, vitae molestie dui bibendum sit amet. Donec mattis mollis nisl, nec mattis ipsum cursus nec. Cras interdum vehicula turpis ut congue. Vestibulum consequat sem ut dui vulputate sed scelerisque justo pulvinar. Praesent ut congue tortor. Suspendisse euismod feugiat fermentum. Sed non magna ut odio egestas molestie ut a velit. Duis interdum auctor mi, a ornare justo malesuada a.
+
'''7/9/2010'''
 +
*minipreps of luxCDE: samples 1&4 usable
-
Nunc laoreet eleifend ultricies. Aenean vestibulum nisi vel justo blandit id commodo nunc luctus. Pellentesque vitae nunc lorem. Sed vehicula consectetur neque ac convallis. Fusce mattis ligula sit amet sem vehicula aliquet. Duis pulvinar laoreet neque eget egestas. Integer eu justo sapien. Vivamus purus lacus, vehicula id ullamcorper at, tempus nec velit. Curabitur vel odio turpis. Donec iaculis rhoncus porttitor. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Nunc magna lectus, ornare nec venenatis nec, interdum at nunc.
+
{| 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