Team:Bordeaux/NotebookJune19

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(Created page with "{{:Team:Bordeaux/Tete}} <html> <h1>19<sup>th</sup> June </h1> <div class="type-exp"> Molecular biology</div> <br> <ul class="day-detail"> <li>PCR of our inserts 40 and 60 </li...")
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<li>PCR of our inserts 40 and 60 </li>
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<li>PCR of our inserts 40 and 60 </li><br>
<li>Digestion of psB1C3 and our inserts 20-40-60 <BR><br>
<li>Digestion of psB1C3 and our inserts 20-40-60 <BR><br>
Concentration PCR 40 = 321,3ng/µl<BR>
Concentration PCR 40 = 321,3ng/µl<BR>
  PCR 60 = 521,0ng/µl<br>
  PCR 60 = 521,0ng/µl<br>
Concentration psB1C3 = 96,5ng/µl -> plate 4 kit 2014<br><br>
Concentration psB1C3 = 96,5ng/µl -> plate 4 kit 2014<br><br>
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digestion psB1C3 : </li>
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Digestion psB1C3 : </li><br>
<table>
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With vector = 5,18µL psB1C3 + 4,82µL H2O
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With vector = 5,18µL psB1C3 + 4,82µL H2O<br><br>
<li>Ligation</li>
<li>Ligation</li>
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So [plasmid] = 0,96ng/µl (via nanodrop)<br>
So [plasmid] = 0,96ng/µl (via nanodrop)<br>
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  <div style="margin-left:40px ;>  = 0,017 pmol/µl (via the website) </div><br>
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  <div style="margin-left:60px ;>  = 0,017 pmol/µl (via the website) <br>
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          <div style="margin-left:40px ;>  = 0,034 pmol/ 2µl </div><br>
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          = 0,034 pmol/ 2µl </div><br>
So we need : 0,017*3=0,051pmol/µl  0,017 * 6 = 0,102pmol/ 2µl
So we need : 0,017*3=0,051pmol/µl  0,017 * 6 = 0,102pmol/ 2µl

Revision as of 22:16, 17 October 2014

19th June

Molecular biology

  • PCR of our inserts 40 and 60

  • Digestion of psB1C3 and our inserts 20-40-60

    Concentration PCR 40 = 321,3ng/µl
    PCR 60 = 521,0ng/µl
    Concentration psB1C3 = 96,5ng/µl -> plate 4 kit 2014

    Digestion psB1C3 :

    • H2O (qsp 20µl) 7,5µl
    Buffer 2 2µl
    XbaI 0,25µl
    SpeI 0,25µl
    Vector 10µL
    With vector = 5,18µL psB1C3 + 4,82µL H2O

  • Ligation
    • We need to have a ratio [insert] = 3*[plasmid]
    We use the website promega
    • psB1C3 = 2270pb
    • VPGXG 20 = 436pb
    • VPGXG 40 = 736pb
    • VPGXG 60 = 1036pb


    So [plasmid] = 0,96ng/µl (via nanodrop)
    Purification

    • SDS-PAGE Tris-Glycine


    Purification

    • Design of oligo with DNA works (website)

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