Difference between revisions of "Team:Paris Bettencourt/Notebook/Riboswitch"

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<div id="notebookMenu">
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  <ul>
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    <li><a href="#july">July</a></li>
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    <li><a href="#august">August</a></li>
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    <li><a href="#september">September</a></li>
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<a name="july" class="anchor"><h1></h1></a>
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<h1 class="date one">July 27th</h1>
  
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<h2>Riboswitch</h2>
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<h2>Goal</h2>
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E coli DH5-alpha transformation with RFP from iGEM plate 4 - 2H (pSB1C3 backbone)
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<h2>Procedure</h2>
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<ol>
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<li> I took iGEM plate 4 - 2H (RFP, BBa_J04450 in PSB1A3 backbone) and resuspend it in 10 uL of distilled water. </li>
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<li> Used E coli competent cells from NEB 2 tubes c2987 (50uL) : 1 control (C) and one for transformation (T) </li>
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<li> Introduced 2 uL (200-300pg/ul) of my suspended plasmid solution in T.</li>
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<li> Waiting 30 min in ice
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<li> Heat shock 30 sec 42°C </li>
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<li> Then 5min in ice </li>
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<li> Add 200uL SOC medium</li>
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<li>Recovery 2 hours then plate it on 3 different 100ug/mL Ampicillin LB</li>
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<li> Incubate 37°C till tomorrow </li>
  
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<b>Thursday, Jul. 23</b>
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<h2>Results</h2>
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The day after I had colonies growing, nothing on my control
 
<ul>
 
<ul>
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<li>Understood the project, just have to do it.</li>
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I took 2 different colonies and do a 8 hours culture in two different tubes T1 an T2
 
</ul>
 
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<b>Friday, Jul. 24</b>
 
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<ul>
 
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<li>Made the project, it is awesome!</li>
 
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Revision as of 08:22, 1 September 2015

July 27th

Goal

E coli DH5-alpha transformation with RFP from iGEM plate 4 - 2H (pSB1C3 backbone)

Procedure

  1. I took iGEM plate 4 - 2H (RFP, BBa_J04450 in PSB1A3 backbone) and resuspend it in 10 uL of distilled water.
  2. Used E coli competent cells from NEB 2 tubes c2987 (50uL) : 1 control (C) and one for transformation (T)
  3. Introduced 2 uL (200-300pg/ul) of my suspended plasmid solution in T.
  4. Waiting 30 min in ice
  5. Heat shock 30 sec 42°C
  6. Then 5min in ice
  7. Add 200uL SOC medium
  8. Recovery 2 hours then plate it on 3 different 100ug/mL Ampicillin LB
  9. Incubate 37°C till tomorrow

    10. Results

    The day after I had colonies growing, nothing on my control
      I took 2 different colonies and do a 8 hours culture in two different tubes T1 an T2