Team:Heidelberg/Notebook/Sensing Group/Notebook/3rdweek

From 2008.igem.org

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</li>
</li>
<li>
<li>
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<a href="https://2008.igem.org/Team:Heidelberg/Chemotaxis" style="color: white">Modeling<!--[if gt IE 6]><!--></a><!--<![endif]-->
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<a href="https://2008.igem.org/Team:Heidelberg/Modeling" style="color: white">Modeling<!--[if gt IE 6]><!--></a><!--<![endif]-->
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</li>
</li>
<li>
<li>
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<a href="https://2008.igem.org/Team:Heidelberg/Notebook/Sensing_Group/Notebook" style="color: white">Notebook<!--[if gt IE 6]><!--></a><!--<![endif]-->
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<a href="https://2008.igem.org/Team:Heidelberg/Notebook/Overview" style="color: white">Notebook<!--[if gt IE 6]><!--></a><!--<![endif]-->
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<li style="width: 160px">
<li style="width: 160px">
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<a href="https://2008.igem.org/Team:Heidelberg/Project/General_information" style="color: white">Human Practice<!--[if gt IE 6]><!--></a><!--<![endif]-->
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<a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Project_Overview" style="color: white">Human Practice<!--[if gt IE 6]><!--></a><!--<![endif]-->
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                               <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Surveys"><span><span>Surveys</span></span></a></li>
                               <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Surveys"><span><span>Surveys</span></span></a></li>
                               <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Open_Day"><span><span>Open Day</span></span></a></li>
                               <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Open_Day"><span><span>Open Day</span></span></a></li>
-
                               <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Nobel_Prize"><span><span>Nobel Prize</span></span></a></li>
+
                               <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Nobel_Prize"><span><span>Nobel Prize</span></span></a></li>
 +
</ul>
</ul>
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== Monday, 08/18/2008 ==
== Monday, 08/18/2008 ==
-
* PCR for LuxQ-1 and LuxQ-2 for Fusion constructs. Different Mg concentrations between 1-4 % were used. For LuxQ-2 another PCR was run using as template ''V. harveyi'' colonies
+
* PCR again for LuxQ-1 and LuxQ-2 for Fusion constructs. Different Mg concentrations from 1% to 4% were used. For LuxQ-2 another PCR was run using as template ''V. harveyi'' colonies
[[Image:HD 080818-F1 LuxQ1 2.png|center|thumb|500px|PCR of LuxQ-1 and LuxQ-2 for subsequent fusion PCR]]
[[Image:HD 080818-F1 LuxQ1 2.png|center|thumb|500px|PCR of LuxQ-1 and LuxQ-2 for subsequent fusion PCR]]
-
* Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar, 1 µl LuxQ)
+
* Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar mixed with 1 µl LuxQ)
* preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep
* preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep
-
* digestion of pDK48 with NcoI/NdeI  
+
* digestion of pDK48 with NcoI/NdeI (NEBuffer 4)
-
* digestion of pTrc99alpha and LuxQ (with BamHI and NcoI) and gelextraction
+
* digestion of pTrc99alpha and LuxQ with BamHI/NcoI (NEBuffer 3 + BSA) and gel extraction
== Tuesday, 08/19/2008 ==
== Tuesday, 08/19/2008 ==
* Miniprep of pDK48 and LuxS
* Miniprep of pDK48 and LuxS
* gel of Fusion constructs and gelextraction of pDK48 from yesterday
* gel of Fusion constructs and gelextraction of pDK48 from yesterday
-
* afterwards another gel was run to purify the Fusion constructs via gelextraction
+
* afterwards another gel was run to purify (get rid of the primers used) the Fusion constructs via gel extraction
-
[[Image:HD 080819-Fusion PCR pDK48-Digestion.png|center|thumb|500px|Fusion PCR and pDK48 gel extraction]]
+
[[Image:HD 080819-Fusion PCR pDK48-Digestion.png|center|thumb|500px|Fusion constructs and pDK48 gel extraction]]
== Wednesday, 08/20/2008 - Friday, 08/22/2008 ==
== Wednesday, 08/20/2008 - Friday, 08/22/2008 ==

Latest revision as of 15:36, 27 October 2008

Back to the overview

Monday, 08/18/2008

  • PCR again for LuxQ-1 and LuxQ-2 for Fusion constructs. Different Mg concentrations from 1% to 4% were used. For LuxQ-2 another PCR was run using as template V. harveyi colonies
PCR of LuxQ-1 and LuxQ-2 for subsequent fusion PCR
  • Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar mixed with 1 µl LuxQ)
  • preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep
  • digestion of pDK48 with NcoI/NdeI (NEBuffer 4)
  • digestion of pTrc99alpha and LuxQ with BamHI/NcoI (NEBuffer 3 + BSA) and gel extraction

Tuesday, 08/19/2008

  • Miniprep of pDK48 and LuxS
  • gel of Fusion constructs and gelextraction of pDK48 from yesterday
  • afterwards another gel was run to purify (get rid of the primers used) the Fusion constructs via gel extraction
Fusion constructs and pDK48 gel extraction

Wednesday, 08/20/2008 - Friday, 08/22/2008

  • nothing important to report

Go to 4th week

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