Team:Toulouse/Notebook/project-monitoring

From 2014.igem.org

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<div class="thelanguage">
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<p class="title2">1. Amplification of Binding Module (pEX-K4) into E.coli</p>
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<p class="title2">1. Amplification of Binding Module (pEX-K4) into <i>E. coli</i></p>
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<p class="title3">Transformation of Binding module (pEX-K4) into E. coli</p>
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<p class="title3">Transformation of Binding module (pEX-K4) into <I>E. coli</i></p>
<p class="texte">Gene "Binding module" synthesized by Eurofins, resuspended in 20μL Tris 10mM
<p class="texte">Gene "Binding module" synthesized by Eurofins, resuspended in 20μL Tris 10mM
<br><b>Result:</b>  We obtained distinct colonies on plates LB + Kanamycin (50 µg/mL)</p>
<br><b>Result:</b>  We obtained distinct colonies on plates LB + Kanamycin (50 µg/mL)</p>
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<p class="title3">Liquid culture of 2 clones : 1 et 2 (pEX-K4) transformed into E. coli</p>
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<p class="title3">Liquid culture of two clones: 1 et 2 (pEX-K4) transformed into <i>E. coli</i></p>
<p class="texte"><b>Date:</b> 01/08/2014</p>
<p class="texte"><b>Date:</b> 01/08/2014</p>
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<p class="title3">Miniprep with QIAprep Spin Miniprep Kit: 2 clones of Binding Module (pEX-K4) into E. coli</p>
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<p class="title3">Miniprep with QIAprep Spin Miniprep Kit: two clones of Binding Module (pEX-K4) into <i>E. coli</i></p>
<p class="texte"><b>Date:</b> 04/08/2014
<p class="texte"><b>Date:</b> 04/08/2014
<br><b>Result:</b> Binding Module (pEX-K4) obtained</p>
<br><b>Result:</b> Binding Module (pEX-K4) obtained</p>
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<p class="texte"><b>Date:</b> 04/08/2014
<p class="texte"><b>Date:</b> 04/08/2014
<br><b>Result:</b>  
<br><b>Result:</b>  
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<br>- 3 bands : 1500bp (vector with Kanamycin resistance), 1300bp (Module Binding) and 1000p bp (vector with pUC Ori)
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<br>- 3 bands : 1500bp (vector with Kanamycin resistance), 1300bp (Module Binding) and 1000bp (vector with pUC Ori)
<br>- The two Binding Module clones are ok</p>
<br>- The two Binding Module clones are ok</p>
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<p class="title2">2. Cloning Binding Module on pEX-K4 with pSB1C3 (BBa_K606013 without RFP) into E. coli</p>
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<p class="title2">2. Cloning Binding Module on pEX-K4 with pSB1C3 (<a href="http://parts.igem.org/Part:BBa_K606013">BBa_K606013</a> without RFP) into <i>E. coli</i></p>
<p class="title3">Digestion Binding Module on pEX-K4 and BBa_K606013</p>
<p class="title3">Digestion Binding Module on pEX-K4 and BBa_K606013</p>
<p class="texte"><b>Date:</b> 23/08/2014
<p class="texte"><b>Date:</b> 23/08/2014
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<br><b>Expected bands after digestion for:</b>
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<br><b>Expected bands after digestion:</b>
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<br>- BBa_K606013 : 860 bp for RFP and 2100 bp for vector  pSB1C3
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<br>- BBa_K606013: 860 bp for RFP and 2100 bp for vector  pSB1C3
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<br>- Binding Module : 1400 bp for Binding Module and 1500bp+1000bp for vector pEX_K4
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<br>- Binding Module: 1400 bp for Binding Module and 1500bp + 1000bp for vector pEX_K4
<br>We decide to do a ligation between pSB1C3 and Binding Module without gel cutting and purification but after a PCR clean-up (Thermo scientific, GeneJET PCR purifiction kit #K0701) to remove the enzymes.</p>
<br>We decide to do a ligation between pSB1C3 and Binding Module without gel cutting and purification but after a PCR clean-up (Thermo scientific, GeneJET PCR purifiction kit #K0701) to remove the enzymes.</p>
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<p class="texte"><b>Date:</b> 04/08/2014
<p class="texte"><b>Date:</b> 04/08/2014
<br>Transformation with 10 µL of the ligation mix and plate on Chloremphenicol LA plate
<br>Transformation with 10 µL of the ligation mix and plate on Chloremphenicol LA plate
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<br>Result : many wrong clones</p>
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<br><b>Result:</b> many wrong clones</p>
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<p class="title2">3. Cloning Binding Module on pEX-K4 with pVeg on pSB1C3 (BBa_K823003) into E. coli</p>
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<p class="title2">3. Cloning Binding Module on pEX-K4 with pVeg on pSB1C3 (<a href="http://parts.igem.org/Part:BBa_K823003">BBa_K823003</a>) into <i>E. coli</i></p>
<p class="title3">Digestion Binding Module on pEX-K4 and BBa_K823003</p>
<p class="title3">Digestion Binding Module on pEX-K4 and BBa_K823003</p>
<p class="texte"><b>Date:</b> 04/08/2014
<p class="texte"><b>Date:</b> 04/08/2014
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<br>Gel Electrophoresis
<br>Gel Electrophoresis
<br><b>Result:</b>
<br><b>Result:</b>
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<br>- Binding Module : 1400 bp for Binding Module and 1500bp+1000bp for vector pEX_K47
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<br>- Binding Module : 1400 bp for Binding Module and 1500bp + 1000bp for vector pEX_K47
<br>We decide to do a ligation between pVeg and Binding Module without gel cutting and purification but after a PCR clean-up (Thermo scientific, GeneJET PCR purifiction kit #K0701) to remove the enzymes.</p>
<br>We decide to do a ligation between pVeg and Binding Module without gel cutting and purification but after a PCR clean-up (Thermo scientific, GeneJET PCR purifiction kit #K0701) to remove the enzymes.</p>
<p class="title3">Ligation Binding Module on pVeg with pSB1C3</p>
<p class="title3">Ligation Binding Module on pVeg with pSB1C3</p>
<p class="texte"><b>Date:</b> 04/08/2014
<p class="texte"><b>Date:</b> 04/08/2014
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<br>BBa_K823003 digested by XbaI and PstI and Binding Module digested by SpeIand PstI and ligation</p>
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<br>BBa_K823003 digested by XbaI and PstI and Binding Module digested by SpeI and PstI and ligation</p>
<p class="title3">Transformation of Binding Module on pVeg into <i>E. coli</i></p>
<p class="title3">Transformation of Binding Module on pVeg into <i>E. coli</i></p>
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<br><b>Result:</b> Many good clones (check on 06/08/2014)</p>
<br><b>Result:</b> Many good clones (check on 06/08/2014)</p>
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<p class="title2">4. Cloning Binding Module with Pveg (BBa-K1364005) on pSBBS4S (BBa-K823022) into <i>E. coli</i></p>
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<p class="title2">4. Cloning Binding Module with P<sub>veg</sub> (<a href="http://parts.igem.org/Part:BBa_K1364005">BBa_K1364005</a>) on pSB<sub>BS</sub>4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>) into <i>E. coli</i></p>
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<p class="title3">Digestion Binding Module with Pveg (BBa-K1364005) and pSBBS4S (BBa-K823022)</p>
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<p class="title3">Digestion Binding Module with P<sub>veg<sub> (<a href="http://parts.igem.org/Part:BBa_K1364005">BBa_K1364005</a>) and pSB<sub>BS</sub>4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>)</p>
<p class="texte"><b>Date:</b> 07/08/2014
<p class="texte"><b>Date:</b> 07/08/2014
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<br>BBa_K823022 and Binding Module with Pveg (BBa_K1364005) digested by EcoRIand PstIGel Electrophoresis
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<br>BBa_K823022 and Binding Module with P<sub>veg</sub> (BBa_K1364005) digested by EcoRI and PstI Gel Electrophoresis
<br><b>Result:</b>
<br><b>Result:</b>
<br>- Binding Module with Pveg 1600 bp for Binding Module and 2100bp for vector pSB1C3</p>
<br>- Binding Module with Pveg 1600 bp for Binding Module and 2100bp for vector pSB1C3</p>
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<p class="title3">Ligation Binding Module with Pveg on pSBbs4S</p>
<p class="title3">Ligation Binding Module with Pveg on pSBbs4S</p>
<p class="texte"><b>Date:</b> 07/08/2014
<p class="texte"><b>Date:</b> 07/08/2014
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<br>BBa_K823022 and Binding Module with Pveg (BBa_K1364005) digested by EcoRIand PstI
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<br>BBa_K823022 and Binding Module with P<sub>veg</sub> (BBa_K1364005) digested by EcoRI and PstI
<br>Ligation
<br>Ligation
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<br><b>Result:</b> Ligation between Binding Module with Pveg on pSBBS4S</p>
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<br><b>Result:</b> Ligation between Binding Module with P<sub>veg</sub> on pSB<sub>BS</sub>4S</p>
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<p class="title3">Transformation of Binding Module with Pveg on pSBBS4S into <i>E. coli</i></p>
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<p class="title3">Transformation of Binding Module with P<sub>veg</sub> on pSB<sub>BS</sub>4S into <i>E. coli</i></p>
<p class="texte"><b>Date:</b> 04/08/2014
<p class="texte"><b>Date:</b> 04/08/2014
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<br>Binding Module with Pveg on pSBbs4S
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<br>Binding Module with P<sub>veg</sub> on pSB<sub>BS</sub>4S
<br>Plate on Ampicillin LA plate
<br>Plate on Ampicillin LA plate
<br><b>Result:</b> Many good clones (check on 13/08/2014)</p>
<br><b>Result:</b> Many good clones (check on 13/08/2014)</p>
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<p class="title3">Transformation of Binding Module with Pveg on pSBBS4S into <i>B. subtilis</i></p>
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<p class="title3">Transformation of Binding Module with P<sub>veg</sub> on pSB<sub>BS</sub>4S into <i>B. subtilis</i></p>
<p class="texte"><b>Date:</b> 04/08/2014
<p class="texte"><b>Date:</b> 04/08/2014
<br>After 5 hours of incubation and the linearization of 6µl of DNA with 1µl of ScaI, we make the transformation. Plate on Spectinomycin LA plate.
<br>After 5 hours of incubation and the linearization of 6µl of DNA with 1µl of ScaI, we make the transformation. Plate on Spectinomycin LA plate.
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<br>Result : One good clone : PCR (check on 09/09/2014) and threonine test (check on 09/09/2014)</p>
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<br><b>Result:</b> One good clone : PCR (check on 09/09/2014) and threonine test (check on 09/09/2014)</p>
<p class="title2">5. Binding Test</p>
<p class="title2">5. Binding Test</p>

Revision as of 20:08, 16 October 2014