Team:Toulouse/Notebook/project-monitoring

From 2014.igem.org

(Difference between revisions)
 
(7 intermediate revisions not shown)
Line 157: Line 157:
       <div class="centering" style="padding-top: 85px; padding-bottom:40px;">
       <div class="centering" style="padding-top: 85px; padding-bottom:40px;">
-
<p style="font-size:1.3em; margin: 0 0 30px 0;"><a href="#" onClick="ddaccordion.collapseall('technology'); return false">Collapse all</a> | <a href="#" onClick="ddaccordion.expandall('technology'); return false">Expand all</a></p>
+
<p style="font-size:1.3em; margin: 0 0 30px 0;"><a href="#" onClick="ddaccordion.expandall('technology'); return false">Expand all</a> | <a href="#" onClick="ddaccordion.collapseall('technology'); return false">Collapse all</a></p>
<div class="technology">Binding module</div>
<div class="technology">Binding module</div>
Line 182: Line 182:
<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="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 <a href="http://parts.igem.org/Part:BBa_K606013">BBa_K606013</a></p>
<p class="texte"><b>Date:</b> 23/08/2014
<p class="texte"><b>Date:</b> 23/08/2014
<br><b>Expected bands after digestion:</b>
<br><b>Expected bands after digestion:</b>
-
<br>- BBa_K606013: 860 bp for RFP and 2100 bp for vector  pSB1C3
+
<br>- <a href="http://parts.igem.org/Part:BBa_K606013">BBa_K606013</a>: 860 bp for RFP and 2100 bp for vector  pSB1C3
<br>- Binding Module: 1400 bp for Binding Module and 1500bp + 1000bp for vector pEX_K4
<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>
Line 198: Line 198:
<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="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 <a href="http://parts.igem.org/Part:BBa_K823003">BBa_K823003</a></p>
<p class="texte"><b>Date:</b> 04/08/2014
<p class="texte"><b>Date:</b> 04/08/2014
-
<br>BBa_K823003 digested by XbaI and PstI and Binding Module digested by SpeIand PstI
+
<br><a href="http://parts.igem.org/Part:BBa_K823003">BBa_K823003</a> digested by XbaI and PstI and Binding Module digested by SpeIand PstI
<br>Gel Electrophoresis
<br>Gel Electrophoresis
<br><b>Result:</b>
<br><b>Result:</b>
Line 208: Line 208:
<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
-
<br>BBa_K823003 digested by XbaI and PstI and Binding Module digested by SpeI and PstI and ligation</p>
+
<br><a href="http://parts.igem.org/Part:BBa_K823003">BBa_K823003</a> 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>
Line 218: Line 218:
<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="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
-
<br>BBa_K823022 and Binding Module with P<sub>veg</sub> (BBa_K1364005) digested by EcoRI and PstI Gel Electrophoresis
+
<br><a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and Binding Module with P<sub>veg</sub> (<a href="http://parts.igem.org/Part:BBa_K1364005">BBa_K1364005</a>) 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>
Line 224: Line 224:
<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
-
<br>BBa_K823022 and Binding Module with P<sub>veg</sub> (BBa_K1364005) digested by EcoRI and PstI
+
<br><a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and Binding Module with P<sub>veg</sub> (<a href="http://parts.igem.org/Part:BBa_K1364005">BBa_K1364005</a>) digested by EcoRI and PstI
<br>Ligation
<br>Ligation
<br><b>Result:</b> Ligation between Binding Module with P<sub>veg</sub> on pSB<sub>BS</sub>4S</p>
<br><b>Result:</b> Ligation between Binding Module with P<sub>veg</sub> on pSB<sub>BS</sub>4S</p>
Line 264: Line 264:
<p class="title2">2. Cloning chemotaxis  <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> (chemotaxis_Puc57) with digested pSB1C3 with EcoRI and PstI into <i>E. coli</i></p>
<p class="title2">2. Cloning chemotaxis  <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> (chemotaxis_Puc57) with digested pSB1C3 with EcoRI and PstI into <i>E. coli</i></p>
-
<p class="title3">Digestion BBa_K1364000 (chemotaxis_Puc57) with EcoRI and PstI - PCR kit Clean up</p>
+
<p class="title3">Digestion <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> (chemotaxis_Puc57) with EcoRI and PstI - PCR kit Clean up</p>
<p class="texte"><b>Date:</b> 07/08/2014
<p class="texte"><b>Date:</b> 07/08/2014
-
<br><b>Result:</b> Expected band after digestion for BBa_K1364000: 2300 bp
+
<br><b>Result:</b> Expected band after digestion for <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a>: 2300 bp
<br><b>Problem:</b> We can't distinguish the vector band (2500 bp)</p>
<br><b>Problem:</b> We can't distinguish the vector band (2500 bp)</p>
-
<p class="title3">Gel extraction of BBa_K1364000</p>
+
<p class="title3">Gel extraction of <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a></p>
<p class="texte"><b>Date:</b> 07/08/2014</p>
<p class="texte"><b>Date:</b> 07/08/2014</p>
-
<p class="title3">Ligation BBa_K1364000 and digested PSB1C3 with EcoRI and PstI</p>
+
<p class="title3">Ligation <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> and digested PSB1C3 with EcoRI and PstI</p>
<p class="texte"><b>Date:</b> 08/08/2014</p>
<p class="texte"><b>Date:</b> 08/08/2014</p>
-
<p class="title3">Transformation BBa_K1364000 in <i>E.coli</i></p>
+
<p class="title3">Transformation <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> in <i>E.coli</i></p>
<p class="texte"><b>Date:</b> 08/08/2014
<p class="texte"><b>Date:</b> 08/08/2014
<br><b>Result:</b> We obtained distinct colonies on LA + Cm (15 µg/mL) plates and resuspended 15 colonies in LB+Cm (15 µg/mL)</p>
<br><b>Result:</b> We obtained distinct colonies on LA + Cm (15 µg/mL) plates and resuspended 15 colonies in LB+Cm (15 µg/mL)</p>
Line 286: Line 286:
<p class="texte"><b>Date:</b> 11/08/2014</p>
<p class="texte"><b>Date:</b> 11/08/2014</p>
-
<p class="title3">Digestion BBa_K1364000 on pSB1C3 with EcoRI and PstI</p>
+
<p class="title3">Digestion <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> on pSB1C3 with EcoRI and PstI</p>
<p class="texte"><b>Date:</b> 11/08/2014
<p class="texte"><b>Date:</b> 11/08/2014
<br><b>Result:</b> There is one colony which presents the right construction.</p>
<br><b>Result:</b> There is one colony which presents the right construction.</p>
Line 293: Line 293:
<p class="title2">3. Cloning chemotaxis <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> (chemotaxis_Puc57) with digested <a href="http://parts.igem.org/Part:BBa_K823003">BBa_823003</a> (P<sub>veg</sub>) on pSB1C3 with SpeI and PstI into <i>E. coli</i></p>
<p class="title2">3. Cloning chemotaxis <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> (chemotaxis_Puc57) with digested <a href="http://parts.igem.org/Part:BBa_K823003">BBa_823003</a> (P<sub>veg</sub>) on pSB1C3 with SpeI and PstI into <i>E. coli</i></p>
-
<p class="title3">Ligation BBa_K1364000 and digested BBa_823003 on PsB1C3 with SpeI and PstI</p>
+
<p class="title3">Ligation <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> and digested <a href="http://parts.igem.org/Part:BBa_K823003">BBa_823003</a> on PsB1C3 with SpeI and PstI</p>
<p class="texte"><b>Date</b>: 08/08/2014</p>
<p class="texte"><b>Date</b>: 08/08/2014</p>
-
<p class="title3">Transformation BBa_1364004 in <i>E.coli</i></p>
+
<p class="title3">Transformation <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_1364004</a> in <i>E.coli</i></p>
<p class="texte"><b>Date:</b> 8/08/2014</p>
<p class="texte"><b>Date:</b> 8/08/2014</p>
<p class="title3">Test of sensibility on Ampicillin</p>
<p class="title3">Test of sensibility on Ampicillin</p>
Line 301: Line 301:
<br><b>Result:</b> We can determine which colony is sensible for ampicillin and know which bacterium is carrying the chemotaxis gene. There is one colony which resists on ampicillin.</p>
<br><b>Result:</b> We can determine which colony is sensible for ampicillin and know which bacterium is carrying the chemotaxis gene. There is one colony which resists on ampicillin.</p>
-
<p class="title3">Digestion BBa_K1364004 on pSBC3 with EcoRI and PstI</p>
+
<p class="title3">Digestion <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_1364004</a> on pSBC3 with EcoRI and PstI</p>
<p class="texte"><b>Date:</b> 12/08/2014
<p class="texte"><b>Date:</b> 12/08/2014
<br><b>Result:</b> We did not see any colony with chemotaxis insert.</p>
<br><b>Result:</b> We did not see any colony with chemotaxis insert.</p>
<p class="title2">4.  Cloning chemotaxis  <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> (chemotaxis_Puc57) with digested <a href="http://parts.igem.org/Part:BBa_K823003">BBa_823003</a> (P<sub>veg</sub>) on pSB1C3 with SpeI and PstI into <i>E. coli</i></p>
<p class="title2">4.  Cloning chemotaxis  <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> (chemotaxis_Puc57) with digested <a href="http://parts.igem.org/Part:BBa_K823003">BBa_823003</a> (P<sub>veg</sub>) on pSB1C3 with SpeI and PstI into <i>E. coli</i></p>
-
<p class="title3">Ligation BBa_K1364000 and digested BBa_823003 on PsB1C3 with SpeI and PstI</p>
+
<p class="title3">Ligation <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a> and digested <a href="http://parts.igem.org/Part:BBa_K823003">BBa_823003</a> on PsB1C3 with SpeI and PstI</p>
<p class="texte"><b>Date:</b> 11/08/2014</p>
<p class="texte"><b>Date:</b> 11/08/2014</p>
-
<p class="title3">Transformation BBa_1364004 in <i>E.coli</i></p>
+
<p class="title3">Transformation <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_1364004</a> in <i>E.coli</i></p>
<p class="texte"><b>Date:</b> 11/08/2014</p>
<p class="texte"><b>Date:</b> 11/08/2014</p>
<p class="title3">Test of sensibility on Ampicillin</p>
<p class="title3">Test of sensibility on Ampicillin</p>
Line 314: Line 314:
<br><b>Result:</b> we obtained 4 colonies sensible at Ampicilline</p>
<br><b>Result:</b> we obtained 4 colonies sensible at Ampicilline</p>
-
<p class="title3">Digestion BBa_K1364004 on PsB1C3 with EcoRI and PstI</p>
+
<p class="title3">Digestion <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_1364004</a> on PsB1C3 with EcoRI and PstI</p>
<p class="texte"><b>Date:</b> 18/08/2014</p>
<p class="texte"><b>Date:</b> 18/08/2014</p>
-
<p class="title3">Gel extraction of BBa_K1364000</p>
+
<p class="title3">Gel extraction of <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_1364000</a></p>
<p class="texte"><b>Date:</b> 19/08/2014</p>
<p class="texte"><b>Date:</b> 19/08/2014</p>
<p class="title2">5. Cloning chemotaxis <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_K1364004</a> with digested pSB<sub>BS</sub>4S with EcorI and PstI into <i>E. coli</i></p>
<p class="title2">5. Cloning chemotaxis <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_K1364004</a> with digested pSB<sub>BS</sub>4S with EcorI and PstI into <i>E. coli</i></p>
-
<p class="title3">Ligation BBa_K1364004 digested EcorI and PstI and digested PsB1C3 with EcoRI and PstI</p>
+
<p class="title3">Ligation <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_K1364004</a> digested EcorI and PstI and digested PsB1C3 with EcoRI and PstI</p>
<p class="texte"><b>Date:</b> 19/08/2014</p>
<p class="texte"><b>Date:</b> 19/08/2014</p>
-
<p class="title3">Transformation BBa_1364004 in pSBBS4S in <i>E.coli</i></p>
+
<p class="title3">Transformation <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_K1364004</a> in pSBBS4S in <i>E.coli</i></p>
<p class="texte"><b>Date:</b> 19/08/2014  
<p class="texte"><b>Date:</b> 19/08/2014  
<br><b>Result:</b> We obtained one colony and resuspended it in LB+ Amp</p>
<br><b>Result:</b> We obtained one colony and resuspended it in LB+ Amp</p>
Line 333: Line 333:
<div class="thelanguage">
<div class="thelanguage">
-
<p class="title2">D4E1</p>
+
<div class="technology2">D4E1</div>
 +
<div class="thelanguage2">
<p class="title2">1. Amplification of synthetic gene (D4E1 on pEX-A2)</p>
<p class="title2">1. Amplification of synthetic gene (D4E1 on pEX-A2)</p>
Line 402: Line 403:
<br><b>Result:</b> clones C, D have the expected construction and are placed in cryopreservation.</p>
<br><b>Result:</b> clones C, D have the expected construction and are placed in cryopreservation.</p>
   
   
-
<p class="title2">4. Cloning Pveg+D4E1 on pSBBS4S (K823022) </p>
+
<p class="title2">4. Cloning P<sub>veg</sub> + D4E1 on pSB<sub>BS</sub>4S (K823022) </p>
<p class="texte"><b>Date:</b> 08/13/2014</p>
<p class="texte"><b>Date:</b> 08/13/2014</p>
   
   
-
<p class="title2">5. Cloning Pveg + D4E1 on pSBBS1C lacZ (23)</p>
+
<p class="title2">5. Cloning P<sub>veg</sub> + D4E1 on pSB<sub>BS</sub>1C lacZ (23)</p>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p>
 +
</div>
 +
 
 +
<div class="technology2">GAFP1</div>
 +
<div class="thelanguage2">
-
<p class="title2">GAFP1</p>
 
<p class="title2">1. Amplification of synthetic gene (GAFP1 on pEX-A2)</p>
<p class="title2">1. Amplification of synthetic gene (GAFP1 on pEX-A2)</p>
Line 428: Line 434:
<p class="title2">2. Cloning GAFP1 gene on PSB1C3 = K1364002 in E. coli</p>
<p class="title2">2. Cloning GAFP1 gene on PSB1C3 = K1364002 in E. coli</p>
-
<p class="title3">Digestion GAFP1_pEX-A2 and BBa_K606013 (RFP_pSB1C3)</p>
+
<p class="title3">Digestion GAFP1_pEX-A2 and <a href="http://parts.igem.org/Part:BBa_K606013">BBa_K606013</a> (RFP_pSB1C3)</p>
<p class="texte"><b>Date:</b> 07/23/2014
<p class="texte"><b>Date:</b> 07/23/2014
-
<br><b>Result:</b> BBa_K606013 : 860 bp
+
<br><b>Result:</b> <a href="http://parts.igem.org/Part:BBa_K606013">BBa_K606013</a> : 860 bp
-
<br>We decide to conserve the miniprep B for BBa_K606013</p>
+
<br>We decide to conserve the miniprep B for <a href="http://parts.igem.org/Part:BBa_K606013">BBa_K606013</a></p>
-
<p class="title3">Ligation GAFP1 and BBa_K606013</p>
+
<p class="title3">Ligation GAFP1 and <a href="http://parts.igem.org/Part:BBa_K606013">BBa_K606013</a></p>
<p class="texte"><b>Date:</b> 07/23/2014</p>
<p class="texte"><b>Date:</b> 07/23/2014</p>
Line 512: Line 518:
-
<p class="title2">5.  Cloning Pveg+GAFP1+Ter B0015 (BBa_K1364008) on pSBBS4S (BBa_K823022)</p>
+
<p class="title2">5.  Cloning Pveg+GAFP1+Ter B0015 (<a href="http://parts.igem.org/Part:BBa_K1364008">BBa_K1364008</a>) on pSBBS4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>)</p>
-
<p class="title3">Digestion of Pveg+GAFP1+ ter B0015 (K1364008) on pSB1C3 and PsBBs4S (K823022)</p>
+
<p class="title3">Digestion of Pveg+GAFP1+ ter B0015 (<a href="http://parts.igem.org/Part:BBa_K1364008">BBa_K1364008</a>) on pSB1C3 and PsBBs4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>)</p>
<p class="texte"><b>Date:</b> 08/01/2014</p>
<p class="texte"><b>Date:</b> 08/01/2014</p>
Line 533: Line 539:
<br><b>Result:</b> clones A, E, F have the right construction</p>
<br><b>Result:</b> clones A, E, F have the right construction</p>
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p>
 +
</div>
 +
<div class="technology2">EcAMP</div>
 +
<div class="thelanguage2">
 +
<p class="texte">The EcAMP part was sent by the Utah iGEM team. It was on a pUC plasmid (ampicilin resistance) with BioBrick suffix and prefix.</p>
-
<p class="title2">EcAMP</p>
+
<p class="title2">1. Transformation of EcAMP in <i>Escherichia coli</i> MC 1061</p>
-
 
+
-
<p class="texte">The EcAMP part was sent by the Utah iGEM team. It was on a pUC plasmid (ampicilin resistance) with biobrick suffix and prefix.</p>
+
-
 
+
-
<p class="title2">1. Transformation of EcAMP in Escherichia coli MC 1061</p>
+
<p class="texte"><b>Date:</b> 07/25/2014</p>
<p class="texte"><b>Date:</b> 07/25/2014</p>
-
<p class="title2">2. Spreading of coli cells transformed with pUC + Utah </p>
+
<p class="title2">2. Spreading of coli cells transformed with EcAMP (plasmid pUC) </p>
<p class="texte"><b>Date:</b> 07/28/2014</p>
<p class="texte"><b>Date:</b> 07/28/2014</p>
-
<p class="title2">3. Liquid culture + Miniprep + Test of the miniprep</p>
+
<p class="title2">3. Liquid culture, Miniprep and Test of the miniprep</p>
<p class="texte"><b>Date:</b> 07/30/2014</p>
<p class="texte"><b>Date:</b> 07/30/2014</p>
-
<p class="title2">4. Cloning 1: EcAMP + Pveg + RBS</p>
+
<p class="title2">4. Cloning 1: EcAMP + P<sub>veg</sub> + RBS</p>
-
<p class="title3">Digestion of EcAMP (INSERT) by XbaI and PstI</p>
+
<p class="title3">Digestion of EcAMP (insert) by XbaI and PstI</p>
<p class="texte"><b>Date:</b> 07/31/2014</p>
<p class="texte"><b>Date:</b> 07/31/2014</p>
-
<p class="title3">Digestion of Pveg + RBS (VECTOR)</p>
+
<p class="title3">Digestion of P<sup>veg</sup> + RBS (VECTOR)</p>
<p class="texte"><b>Date:</b> 07/31/2014</p>
<p class="texte"><b>Date:</b> 07/31/2014</p>
Line 568: Line 575:
<p class="texte"><b>Date:</b> 08/05/2014
<p class="texte"><b>Date:</b> 08/05/2014
-
<br>The first cloning show a lack of DNA in the Miniprep EcAMP. The bands are hardly visible on the gel for the linearization of EcAMP + Pveg + RBS. The problem came from the PCR cleanup step: the fragment size of EcAMP is lower than 200bp.</p>
+
<br><b>Result:</b> The first cloning show a lack of DNA in the Miniprep EcAMP. The bands are hardly visible on the gel for the linearization of EcAMP + Pveg + RBS. The problem came from the PCR cleanup step: the fragment size of EcAMP is lower than 200bp.</p>
<p class="title2">5. Cloning 2: EcAMP + Pveg + RBS </p>
<p class="title2">5. Cloning 2: EcAMP + Pveg + RBS </p>
-
<p class="texte"><b>Date:</b> 06/08/2014</p>
+
<p class="texte"><b>Date:</b> 08/06/2014</p>
-
<p class="title3">Digestion of EcAMP (INSERT) by XbaI and PstI</p>
+
<p class="title3">Digestion of EcAMP (insert) by XbaI and PstI</p>
-
<p class="title3">Digestion of Pveg + RBS (VECTOR)</p>
+
<p class="texte"><b>Date:</b> 08/06/2014</p>
 +
<p class="title3">Digestion of P<sub>veg</sub> + RBS (vector)</p>
 +
<p class="texte"><b>Date:</b> 08/06/2014</p>
<p class="title3">Heat inactivation of the enzymes</p>
<p class="title3">Heat inactivation of the enzymes</p>
 +
<p class="texte"><b>Date:</b> 08/06/2014</p>
<p class="title3">Ligation and transformation</p>
<p class="title3">Ligation and transformation</p>
 +
<p class="texte"><b>Date:</b> 08/06/2014</p>
<p class="title3">PCR test</p>
<p class="title3">PCR test</p>
-
<p class="texte"><b>Date:</b> 07/08/2014</p>
+
<p class="texte"><b>Date:</b> 08/07/2014</p>
<p class="title3">Striation on a petri dish to purify the clone</p>
<p class="title3">Striation on a petri dish to purify the clone</p>
-
<p class="texte">Purpose: to isolate a clone with vector+insert
+
<p class="texte"><b>Purpose:</b> to isolate a clone with vector+insert
<br><b>Date:</b> 08/072014</p>
<br><b>Date:</b> 08/072014</p>
-
<p class="title3">Miniprep of Pveg+SpoVG+EcAMP and analytic digestion</p>
+
<p class="title3">Miniprep of P<sub>veg</sub> + SpoVG + EcAMP and analytic digestion</p>
<p class="texte"><b>Date:</b> 08/08/2014</p>
<p class="texte"><b>Date:</b> 08/08/2014</p>
-
<p class="title3">Ligation of Pveg SpoVG EcAMP with double terminateur B0015 +Transformation and liquid culture</p>
+
<p class="title3">Ligation of Pveg SpoVG EcAMP with double terminateur B0015 + transformation and liquid culture</p>
<p class="texte"><b>Date:</b> 08/11/2014</p>
<p class="texte"><b>Date:</b> 08/11/2014</p>
-
<p class="title3">Miniprep of Pveg SpoVG EcAMP + analytic digestion</p>
+
<p class="title3">Miniprep of P<sub>veg</sub> SpoVG EcAMP + analytic digestion</p>
<p class="texte"><b>Date:</b> 08/13/2014</p>
<p class="texte"><b>Date:</b> 08/13/2014</p>
-
<p class="title3">Cloning K1364011 (EcAMP+Pveg +SpoVG + B0015) + K823022 (psBbs4S) : Digestion, ligation, transformation</p>
+
<p class="title3">Cloning K1364011 (EcAMP + P<sub>veg</sub> +SpoVG + B0015) + K823022 (pSB<sub>BS</sub>4S): Digestion, ligation, transformation</p>
<p class="texte"><b>Date:</b> 08/19/2014</p>
<p class="texte"><b>Date:</b> 08/19/2014</p>
-
<p class="title3">Cloning K1364011 + K823023 (pSBBS1C) : Digestion, ligation, transformation</p>
+
<p class="title3">Cloning K1364011 + K823023 (pSB<sub>BS</sub>1C) : Digestion, ligation, transformation</p>
<p class="texte"><b>Date:</b> 08/18/2014</p>
<p class="texte"><b>Date:</b> 08/18/2014</p>
-
<p class="title3">Verification of the insertion of K1364011 + K823022 (pSBBS4S) into the subtilis genome by threonine test </p>
+
<p class="title3">Verification of the insertion of K1364011 + K823022 (pSB<sub>BS</sub>4S) into the subtilis genome by threonine test </p>
-
<p class="texte"><b>Date:</b> 08/21/2014
+
<p class="texte"><b>Date:</b> 08/21/2014</p>
-
 
+
-
<br>
+
-
<br>Assembling the fungicides module : </p>
+
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p>
 +
</div>
 +
<div class="technology2">Assembling the fungicides module: D4E1 + GAFP1</div>
 +
<div class="thelanguage2">
-
<p class="title2">D4E1 + GAFP1 </p>
+
<p class="title2">1.  Cloning GAFP1+D4E1 on pSB1C3: <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a></p>
-
<p class="title2">1.  Cloning GAFP1+D4E1 on pSB1C3: BBa_K1364012 </p>
+
<p class="title3">Digestion of D4E1 on pEX-A2 and GAFP1 on pSB1C3</p>
<p class="title3">Digestion of D4E1 on pEX-A2 and GAFP1 on pSB1C3</p>
Line 622: Line 633:
<p class="texte"><b>Date:</b> 07/28/2014</p>
<p class="texte"><b>Date:</b> 07/28/2014</p>
   
   
-
<p class="title3">PCR of GAFP1+D4E1 (pSB1C3) A, B, C, D, E, F + electrophoresis</p>
+
<p class="title3">PCR of GAFP1 + D4E1 (pSB1C3) A, B, C, D, E, F + electrophoresis</p>
<p class="texte"><b>Date:</b> 07/29/2014
<p class="texte"><b>Date:</b> 07/29/2014
<br><b>Result:</b> clones A, C, F, G seem to have the expected construction.</p>
<br><b>Result:</b> clones A, C, F, G seem to have the expected construction.</p>
   
   
-
<p class="title3">Digestion of GAFP1+D4E1 (pSB1C3) A, C, F, G + electrophoresis</p>
+
<p class="title3">Digestion of GAFP1 + D4E1 (pSB1C3) A, C, F, G + electrophoresis</p>
<p class="texte"><b>Date:</b> 07/30/2014
<p class="texte"><b>Date:</b> 07/30/2014
-
<br><b>Result:</b> clone  F (BBa_K1364012) has the expected construction and is placed on cryopreservation.</p>
+
<br><b>Result:</b> clone  F (<a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a>) has the expected construction and is placed on cryopreservation.</p>
   
   
-
<p class="title2">2. Cloning GAFP1+D4E1 (BBa_K1364012) on Pveg plasmid (BBa_K823003): BBa_K1364013</p>
+
<p class="title2">2. Cloning GAFP1 + D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a>) on P<sub>veg</sub> plasmid (<a href="http://parts.igem.org/Part:BBa_K823003">BBa_K823003</a>): <a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a></p>
-
<p class="title2">3. Cloning Pveg+GAFP1+D4E1 (BBa_K1364013) on pSBBS4S (K823022)</p>
+
<p class="texte"><b>Date:</b> 08/04/2014
-
<p class="title2">4. Cloning Pveg+GAFP1+D4E1 (BBa_K1364013) on pSBBS1C lacZ (K823023)</p>
+
-
<p class="title2">5. Fungicides tests</p>
+
 +
<p class="title2">3. Cloning P<sub>veg</sub> + GAFP1 + D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a>) on pSB<sub>BS<sub>4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>)</p>
 +
<p class="texte"><b>Date:</b> 08/06/2014
 +
 +
<p class="title2">4. Cloning P<sub>veg</sub> + GAFP1 + D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a>) on pSB<sub>BS</sub>1C lacZ (<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>)</p>
 +
<p class="texte"><b>Date:</b> 08/11/2014
 +
 +
<p class="title2">5. Fungicides tests</p>
 +
<p class="texte"><b>Date:</b> 08/15/2014
-
<p class="title2">Cloning D4E1-GAFP1-EcAMP: BBa_K1364014</p>
+
<p class="title2">Cloning D4E1-GAFP1-EcAMP: <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a></p>
-
<p class="title2">1.  Construction of BBa_K1364014 in PsB1C3, in E. coli</p>
+
<p class="title2">1.  Construction of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> in pSB1C3, in <i>E. coli</i></p>
-
<p class="title3">Digestion of K1364010 and K1364012</p>
+
<p class="title3">Digestion of <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a> and <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a></p>
-
<p class="texte">Digestion of K13664010 by Spe1 and Pst1, and digestion of K1364012 by XbaI and Pst1.
+
<p class="texte">Digestion of <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a> by SpeI and PstI, and digestion of <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a> by XbaI and PstI.
<br><b>Date:</b> 08/11/2014
<br><b>Date:</b> 08/11/2014
-
<br><b>Result:</b> We obtained digested fragments of K1364012 and ~500 bp fragment of K1364010</p>
+
<br><b>Result:</b> We obtained digested fragments of <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a> and ~500 bp fragment of <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a></p>
-
<p class="title3">Ligation of ~500 bp fragment from K1364010 and K1364012</p>
+
<p class="title3">Ligation of ~500 bp fragment from <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a> and <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a></p>
<p class="texte"><b>Date:</b> 08/11/2014
<p class="texte"><b>Date:</b> 08/11/2014
-
<br><b>Result:</b> We obtained ligation of K1364012 and ~500 bp fragment of K1364010</p>
+
<br><b>Result:</b> We obtained ligation of K1364012 and ~500 bp fragment of <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a></p>
-
<p class="title3">Transformation of ligation of ~500 bp fragment from K1364010 and K1364012 = K1364014 in <i>E.coli</i></p>
+
<p class="title3">Transformation of ligation of ~500 bp fragment from <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a> and <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a> = <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> in <i>E.coli</i></p>
<p class="texte"><b>Date:</b> 08/12/2014</p>
<p class="texte"><b>Date:</b> 08/12/2014</p>
-
<p class="title3">Testing 8 <i>E.coli</i> + K1364014 clones</p>
+
<p class="title3">Testing 8 <i>E.coli</i> + <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> clones</p>
<p class="texte"><b>Date:</b> 08/14/2014 </p>
<p class="texte"><b>Date:</b> 08/14/2014 </p>
-
<p class="title3">Testing 15 <i>E.coli</i> + K1364014 clones</p>
+
<p class="title3">Testing 15 <i>E.coli</i> + <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> clones</p>
<p class="texte"><b>Date:</b> 08/18/2014
<p class="texte"><b>Date:</b> 08/18/2014
-
<br><b>Result:</b> clones H, J, O, Q, U, V might have the right K1364014 construction</p>
+
<br><b>Result:</b> clones H, J, O, Q, U, V might have the right <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> construction</p>
-
<p class="title2">2. BBa_K1364014 in PsBs4s (K823022) and PsBs1C (K823023)</p>
+
<p class="title2">2. <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> in pSB<sub>BS</sub>4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>) and pSB<sub>BS</sub>1C (<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>)</p>
-
<p class="title3">Digestion of K1364014, K823022 and K823023</p>
+
<p class="title3">Digestion of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>, <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a></p>
<p class="texte"><b>Date:</b> 08/22/2014
<p class="texte"><b>Date:</b> 08/22/2014
-
<br><b>Result</b>: digestion of K13664014, K823022 and K823023 by EcoR1 and Pst1 were well performed.</p>
+
<br><b>Result</b>: Digestion of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>, <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> by EcoRI and PstI were well performed.</p>
-
<p class="title3">Ligation of K1364014 with K823022 and K1364014 with K823023</p>
+
<p class="title3">Ligation of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> with <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and K1364014 with K823023</p>
<p class="texte"><b>Date:</b> 08/22/2014
<p class="texte"><b>Date:</b> 08/22/2014
-
<br><b>Result:</b> We obtained 20µL ligation of K1364014 with K823022 and of K1364014 with K823023.
+
<br><b>Result:</b> We obtained 20µL ligation of K1364014 with <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> with <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>.
</p>
</p>
<p class="title3">Transformation of ligations in <i>E. coli</i></p>
<p class="title3">Transformation of ligations in <i>E. coli</i></p>
-
<p class="texte">E. coli transformed by K1364014+K823022 spread on LB+Amp 100µg/mL agar plate</p>
+
<p class="texte"><I>E. coli</I> transformed by <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> + <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> spread on LB + Amp 100µg/mL agar plate</p>
-
<p class="texte">E. coli transformed by K1364014+K823023 spread on LB+Amp 100µg/mL agar plate
+
<p class="texte"><i>E. coli</i> transformed by <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> + <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> spread on LB + Amp 100µg/mL agar plate
<br><b>Date:</b> 08/25/2014</p>
<br><b>Date:</b> 08/25/2014</p>
-
<p class="title3">Testing <i>E. coli</i> + K1364014 + K823022 and <i>E. coli </i> + K1364014 + K823023 clones</p>
+
<p class="title3">Testing <i>E. coli</i> + <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> + <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <i>E. coli </i> + <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> + <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> clones</p>
<p class="texte"><b>Date:</b> 08/27/2014</p>
<p class="texte"><b>Date:</b> 08/27/2014</p>
-
<p class="title2">3.  Transformation of K1364014+K823022 and K1364014+K823023 in <i>B. subtilis</i></p>
+
<p class="title2">3.  Transformation of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> + <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> in <i>B. subtilis</i></p>
-
<p class="texte"><i>B. subtilis</i> transformed by 10µL K1364014+K823022 spread on LB+Spec 75µg/mL agar plate
+
<p class="texte"><i>B. subtilis</i> transformed by 10µL <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> spread on LB+Spec 75µg/mL agar plate
-
<br><i>B. subtilis</i> transformed by 10µL K1364014+K823023 spread on LB+Cm 15µg/mL agar plate
+
<br><i>B. subtilis</i> transformed by 10µL <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>  spread on LB+Cm 15µg/mL agar plate
<br><b>Date:</b> 08/28/2014</p>
<br><b>Date:</b> 08/28/2014</p>
-
<p class="title2">Fungicides tests</p>
+
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p>
 +
</div>
 +
 
 +
<div class="technology2">Fungicides tests</div>
 +
<div class="thelanguage2">
 +
 
<p class="title2">1. D4E1-GAFP1</p>
<p class="title2">1. D4E1-GAFP1</p>
-
<p class="title3">Transformation in bacillus  Pveg-D4E1-GAFP1 on pSBBS4S </p>
+
<p class="title3">Transformation of P<sub>veg</sub> - D4E1 - GAFP1 in <i>Bacillus subtilis</i>  on pSB<sub>BS</sub>4S </p>
<p class="texte"><b>Date:</b> 08/12/2014</p>  
<p class="texte"><b>Date:</b> 08/12/2014</p>  
<p class="title3">Integration threonine test + fungicide test </p>
<p class="title3">Integration threonine test + fungicide test </p>
Line 696: Line 718:
<p class="title2"2. D4E1</p>
<p class="title2"2. D4E1</p>
<p class="title3">
<p class="title3">
-
Cloning D4E1 into pSBBS1C + fungicide test</p>
+
Cloning D4E1 into pSB<sub>BS</sub>1C + fungicide test</p>
<p class="texte"><b>Date:</b>08/15/2014</p>
<p class="texte"><b>Date:</b>08/15/2014</p>
<p class="title3">
<p class="title3">
Line 707: Line 729:
<div class="clear"></div>
<div class="clear"></div>
-
 
+
</div>
     </div>
     </div>
   </div>
   </div>

Latest revision as of 01:22, 18 October 2014