Team:Copenhagen/Project/Experimental

From 2011.igem.org

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<table> <!-- Table for content area -->
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   <font color="#000000" face="constantia" size="5">
   <font color="#000000" face="constantia" size="5">
<br>   
<br>   
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<b><center>How to Mutate</center></b><br><br>
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<b><center>Test</center></b><br><br>
   </font>
   </font>
<p align="justify">
<p align="justify">
-
Text.</p>
+
<ul>
 +
<li>Once you have a colony on your plate you take it out and place it in and overnight culture</li>
 +
<li>You perform a Miniprep on your culture to purify your amplified plasmid </li>
 +
<li>Digest your purified plasmid with restrictionsenzymes to check if your plasmids are mutated in the recognition site</li>
 +
<li>Run the digest of the plasmid and hope to see a single band and not two (in the latter case your mutations failed) </li>
 +
<li>Add prefix and suffix containing primers for the CYP in question and amplify with PCR</li>
 +
<li>Purify with DNA purification kit</li>
 +
<li>Cut with restrictionsenzymes EcoR1 and Pst1</li>
 +
<li>Run on a gel and cut the wanted band out </li>
 +
<li>Put it in the freezer</li>
 +
<li>Cut the linarized plasmid backbone with Pst1 and EcoR1 </li>
 +
<li>Ligate CYP and Plasmid </li>
 +
<li>Transform in XL1-Blue</li>
 +
<li>Overnight culture</li>
 +
<li>Hurra - You have a Biobrick</li>
 +
</ul></p>
  </td>
  </td>
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   <td width="250px" height="100%" valign="top">
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   <td width="300px" height="100%" valign="top">
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   <font color="#000000" face="constantia size="7">
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   <font color="#000000" face="constantia" size="5">
<br>   
<br>   
<b><center>How to make a BioBrick</center></b><br><br>
<b><center>How to make a BioBrick</center></b><br><br>
   </font>
   </font>
-
<br>
 
<p align="justify">
<p align="justify">
-
Once you have a colony on your plate you take it out and place it in and overnight culture
+
<ul>
-
<br><br>
+
<li>Once you have a colony on your plate you take it out and place it in and overnight culture</li>
-
You perform a Miniprep on your culture to purify your amplified plasmid  
+
<li>You perform a Miniprep on your culture to purify your amplified plasmid </li>
-
<br><br>
+
<li>Digest your purified plasmid with restrictionsenzymes to check if your plasmids are mutated in the recognition site</li>
-
Digest your purified plasmid with restrictionsenzymes to check if your plasmids are mutated in the recognition site  
+
<li>Run the digest of the plasmid and hope to see a single band and not two (in the latter case your mutations failed) </li>
-
<br><br>
+
<li>Add prefix and suffix containing primers for the CYP in question and amplify with PCR</li>
-
Run the digest of the plasmid and hope to see a single band and not two (in the latter case your mutations failed)
+
<li>Purify with DNA purification kit</li>
-
<br><br>
+
<li>Cut with restrictionsenzymes EcoR1 and Pst1</li>
-
Add prefix and suffix containing primers for the CYP in question and run a PCR  
+
<li>Run on a gel and cut the wanted band out </li>
-
<br><br>
+
<li>Put it in the freezer</li>
-
Purify with DNA purification kit
+
<li>Cut the linarized plasmid backbone with Pst1 and EcoR1 </li>
-
<br><br>
+
<li>Ligate CYP and Plasmid </li>
-
Cut with restrictionsenzymes EcoR1 and Pst1
+
<li>Transform in XL1-Blue</li>
-
<br><br>
+
<li>Overnight culture</li>
-
Run on a gel and cut the wanted band out
+
<li>Hurra - You have a Biobrick</li>
-
<br>
+
</ul></p>
-
http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.pdf
+
-
<br>
+
-
<br>
+
-
Put it in the freezer
+
-
<br><br>
+
-
Cut the linarized plasmid backbone with Pst1 and EcoR1
+
-
<br>http://partsregistry.org/Help:Protocols/Linearized_Plasmid_Backbones
+
-
<br>
+
-
<br>
+
-
Ligate CYP and Plasmid
+
-
<br> http://partsregistry.org/Help:Protocols/Linearized_Plasmid_Backbones
+
-
<br><br>
+
-
Transform in XL1-Blue  
+
-
<br><br> Overnight cultures
+
-
<br><br> Miniprep
+
-
<br><br> Voila - you have a BioBrick
+
-
</p>
+
-
 
+
  </td>
  </td>
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   <td width="250px" height="100%" valign="top">
+
   <td width="300px" height="100%" valign="top">
   <font color="#000000" face="constantia" size="5">
   <font color="#000000" face="constantia" size="5">
<br>   
<br>   
-
<b><center><How to make a CyperMan</center></b><br><br>
+
<b><center>Test</center></b><br><br>
   </font>
   </font>
<p align="justify">
<p align="justify">
-
Text.</p>
+
<ul>
 +
<li>Once you have a colony on your plate you take it out and place it in and overnight culture</li>
 +
<li>You perform a Miniprep on your culture to purify your amplified plasmid </li>
 +
<li>Digest your purified plasmid with restrictionsenzymes to check if your plasmids are mutated in the recognition site</li>
 +
<li>Run the digest of the plasmid and hope to see a single band and not two (in the latter case your mutations failed) </li>
 +
<li>Add prefix and suffix containing primers for the CYP in question and amplify with PCR</li>
 +
<li>Purify with DNA purification kit</li>
 +
<li>Cut with restrictionsenzymes EcoR1 and Pst1</li>
 +
<li>Run on a gel and cut the wanted band out </li>
 +
<li>Put it in the freezer</li>
 +
<li>Cut the linarized plasmid backbone with Pst1 and EcoR1 </li>
 +
<li>Ligate CYP and Plasmid </li>
 +
<li>Transform in XL1-Blue</li>
 +
<li>Overnight culture</li>
 +
<li>Hurra - You have a Biobrick</li>
 +
</ul></p>
  </td>
  </td>

Revision as of 11:58, 11 July 2011


Test

  • Once you have a colony on your plate you take it out and place it in and overnight culture
  • You perform a Miniprep on your culture to purify your amplified plasmid
  • Digest your purified plasmid with restrictionsenzymes to check if your plasmids are mutated in the recognition site
  • Run the digest of the plasmid and hope to see a single band and not two (in the latter case your mutations failed)
  • Add prefix and suffix containing primers for the CYP in question and amplify with PCR
  • Purify with DNA purification kit
  • Cut with restrictionsenzymes EcoR1 and Pst1
  • Run on a gel and cut the wanted band out
  • Put it in the freezer
  • Cut the linarized plasmid backbone with Pst1 and EcoR1
  • Ligate CYP and Plasmid
  • Transform in XL1-Blue
  • Overnight culture
  • Hurra - You have a Biobrick
How to make a BioBrick

  • Once you have a colony on your plate you take it out and place it in and overnight culture
  • You perform a Miniprep on your culture to purify your amplified plasmid
  • Digest your purified plasmid with restrictionsenzymes to check if your plasmids are mutated in the recognition site
  • Run the digest of the plasmid and hope to see a single band and not two (in the latter case your mutations failed)
  • Add prefix and suffix containing primers for the CYP in question and amplify with PCR
  • Purify with DNA purification kit
  • Cut with restrictionsenzymes EcoR1 and Pst1
  • Run on a gel and cut the wanted band out
  • Put it in the freezer
  • Cut the linarized plasmid backbone with Pst1 and EcoR1
  • Ligate CYP and Plasmid
  • Transform in XL1-Blue
  • Overnight culture
  • Hurra - You have a Biobrick
Test

  • Once you have a colony on your plate you take it out and place it in and overnight culture
  • You perform a Miniprep on your culture to purify your amplified plasmid
  • Digest your purified plasmid with restrictionsenzymes to check if your plasmids are mutated in the recognition site
  • Run the digest of the plasmid and hope to see a single band and not two (in the latter case your mutations failed)
  • Add prefix and suffix containing primers for the CYP in question and amplify with PCR
  • Purify with DNA purification kit
  • Cut with restrictionsenzymes EcoR1 and Pst1
  • Run on a gel and cut the wanted band out
  • Put it in the freezer
  • Cut the linarized plasmid backbone with Pst1 and EcoR1
  • Ligate CYP and Plasmid
  • Transform in XL1-Blue
  • Overnight culture
  • Hurra - You have a Biobrick
Comments or questions to the team? Please

Retrieved from "http://2011.igem.org/Team:Copenhagen/Project/Experimental"