Team:EPF-Lausanne/Notebook/Cloning Strategy

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{{EPF-Lausanne09}}
 
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<div CLASS="epfltrick">__TOC__
 
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</div><div CLASS="epfl09">
 
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=Cloning strategy=
 
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===14.07.09===
 
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[[Media: primer15+160709.pdf| Primers designed for LOVTAP read-out and RBphP project.]]
 
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===15.07.09===
 
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===20.07.09===
 
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===21.07.09===
 
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One useful website to know the restriction sites of enzymes:
 
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<br>http://www.genscript.com/cgi-bin/products/enzyme.cgi?op=all_ez
 
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The restriction site used were:
 
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<br>EcoRI          GAATTC
 
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<br>XbaI          TCTAGA
 
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<br>SpeI          ACTAGT
 
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<br>PsiI          TTATAA
 
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Design the primers for the 2 step-PCR:
 
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the first step introduces the LacI promoter and the RBS upstream the LovTAP gene with the Forward primer, whereas the Reverse introduces the Term downstream.
 
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In the second step we only introduce the E-X restriction sites upstream and the SP downstream.
 
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===22.07.09===
 
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===23.07.09===
 
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===24.07.09===
 
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===27.07.09===
 
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===28.07.09===
 
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===29.07.09===
 
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===30.07.09===
 
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===31.07.09===
 
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==August==
 
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</div><div CLASS="epfl09bouchon"></div>
 

Latest revision as of 08:05, 28 July 2009