Team:Bordeaux/NotebookAugust12

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(Created page with "{{:Team:Bordeaux/Tete}} <html> <h1>12<sup>th</sup> August </h1> <div class="type-exp"> Molecular biology</div> <br> <ul class="strain-title"> <li>Digestion We want a DNA conce...")
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<br>Final concentration after digestion :  
<br>Final concentration after digestion :  
<li>Plasmid: 18 ng/μl</li>
<li>Plasmid: 18 ng/μl</li>
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<li>Insert: 20 ng/μl</li>
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<li>Insert: 20 ng/μl</li></ul>
<ul class="day-detail">
<ul class="day-detail">

Revision as of 01:33, 18 October 2014

12th August

Molecular biology

  • Digestion We want a DNA concentration of 100ng/μl
    Time-Saver protocol
    DNA 10μl
    Xba1 0,5μl
    Spe1 0,5μl
    Buffer 10X 5μl
    H2O qsp 50μl
    => 15min at 37°C
    => Inactivation of enzyme 20min at 80°C

    • Final concentration after digestion :
  • Plasmid: 18 ng/μl
  • Insert: 20 ng/μl
  • Ligation
    ELP – 369bp
    SLP – 369bp
    RLP – 357bp
    Plasmid – 2270bp
    Plasmid 4μl
    Insert 1,8μl
    Buffer 10X 1μl
    Taq ligase 0,5μl
    H2O qsp 10μl
  • Making competent cells
    BL21 in LB+Amp plates

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