Difference between revisions of "Team:Brasil-USP/Notebook/Calendar"

Line 605: Line 605:
  
  
−
<a id="August"></a><h1>August</h1>
+
<h1>August</h1>
  
  
Line 639: Line 639:
  
 
<br>       
 
<br>       
−
- EXSP digestion (R0010, R0080, B0034, Pt_I13507)<br><br>   
+
- EXSP digestion (<a href="http://parts.igem.org/Part:BBa_R0010">R0010</a>, <a href="http://parts.igem.org/Part:BBa_R0080">R0080</a>, <a
−
- NotI digestion (Pl_I13507)<br><br>   
+
 
−
- Ligation Pa_RBS (R0080 + B0034) and Pl_RBS (R0010 + B0034)<br><br>
+
href="http://parts.igem.org/Part:BBa_B0034">B0034</a>, <a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pt-I13507.pdf">Pt_I13507</a>)<br><br>   
 +
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>)<br><br>   
 +
- Ligation <a href="https://static.igem.org/mediawiki/2015/9/97/Team-Brasil-USP_Pa-RBS.pdf">Pa_RBS</a> (R0080 + B0034) and <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/c/c6/Team-Brasil-USP_Pl_RBS.pdf">Pl_RBS</a> (R0010 + B0034)<br><br>
 
<br>
 
<br>
 
        
 
        
Line 648: Line 652:
 
<br>
 
<br>
  
−
- NotI digestion (Pr_I13507)<br><br>
+
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>)<br><br>
−
- Agarose gel (Pr_I13507) - Positive results!!<br><br>
+
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>) - Positive results!!<br><br>
−
- EXSP digestion (Pt_I13507)<br><br>
+
- EXSP digestion (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pt-I13507.pdf">Pt_I13507</a>)<br><br>
−
- Transformation (Pl_RBS and Pa_RBS)<br><br>
+
- Transformation (<a href="https://static.igem.org/mediawiki/2015/c/c6/Team-Brasil-USP_Pl_RBS.pdf">Pl_RBS</a> and <a href="https://static.igem.org/mediawiki/2015/9/97/Team-
 +
 
 +
Brasil-USP_Pa-RBS.pdf">Pa_RBS</a>)<br><br>
  
 
<br>
 
<br>
Line 657: Line 663:
 
     <td valign=top align=left> 05/08
 
     <td valign=top align=left> 05/08
 
<br>
 
<br>
−
- agarose gel (Pl_I13507, Pt_I13507)- negative results for Pl_13507 :(<br><br>
+
- agarose gel (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/4/41/Team-
−
- Agarose gel  and gel purification (Pt_I13507)<br><br>
+
 
 +
Brasil-USP_Pt-I13507.pdf">Pt_I13507</a>)- negative results for <a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a> :(<br><br>
 +
- Agarose gel  and gel purification (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pt-I13507.pdf">Pt_I13507</a>)<br><br>
 
- ligation Pl_I13507 (Pl_P0440 + Pt_I13507)<br><br>
 
- ligation Pl_I13507 (Pl_P0440 + Pt_I13507)<br><br>
−
- inoculum (Pl_RBS, Pa_RBS)<br><br>
+
- inoculum (<a href="https://static.igem.org/mediawiki/2015/c/c6/Team-Brasil-USP_Pl_RBS.pdf">Pl_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/9/97/Team-Brasil-USP_Pa-
 +
 
 +
RBS.pdf">Pa_RBS</a>)<br><br>
  
 
<br>
 
<br>
Line 670: Line 680:
 
     <td valign=top align=left> 06/08
 
     <td valign=top align=left> 06/08
 
<br>     
 
<br>     
−
- Transformation (Pl_I13507, Pr_I13507)<br><br>
+
- Transformation (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-
−
- miniprep (Pl_RBS, Pa_RBS)<br><br>
+
 
−
- NotI digestion (Pl_RBS, Pa_RBS)<br><br>
+
Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>)<br><br>
 +
- miniprep (<a href="https://static.igem.org/mediawiki/2015/c/c6/Team-Brasil-USP_Pl_RBS.pdf">Pl_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/9/97/Team-Brasil-USP_Pa-
 +
 
 +
RBS.pdf">Pa_RBS</a>)<br><br>
 +
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/c/c6/Team-Brasil-USP_Pl_RBS.pdf">Pl_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/9/97/Team-Brasil-
 +
 
 +
USP_Pa-RBS.pdf">Pa_RBS</a>)<br><br>
  
 
<br>
 
<br>
Line 679: Line 695:
 
     <td valign=top align=left> 07/08
 
     <td valign=top align=left> 07/08
 
<br>
 
<br>
−
- Agarose gel (Pl_RBS, Pa_RBS) - Only Pa_RBS was positive<br><br>  
+
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/c/c6/Team-Brasil-USP_Pl_RBS.pdf">Pl_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/9/97/Team-Brasil-
−
- EXSP digestion (Pa_RBS, Pt_Term)<br><br>
+
 
−
- Ligation Pa_Term(Pa_RBS + Pt_Term) Pa_I13507 (Pa_RBS + GFP_I13507)<br><br>
+
USP_Pa-RBS.pdf">Pa_RBS</a>) - Only Pa_RBS was positive<br><br>  
 +
- EXSP digestion (<a href="https://static.igem.org/mediawiki/2015/9/97/Team-Brasil-USP_Pa-RBS.pdf">Pa_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-
 +
 
 +
USP_Pt-E0840.pdf">Pt_Term</a>)<br><br>
 +
- Ligation <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-Term.pdf">Pa_Term</a>(Pa_RBS + Pt_Term), <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a> (Pa_RBS + GFP_I13507)<br><br>
  
 
<br>
 
<br>
Line 689: Line 711:
 
<br>
 
<br>
 
- Ligation (2X_RBS_hokD + Term)<br><br>
 
- Ligation (2X_RBS_hokD + Term)<br><br>
−
- Transformation (Pa_I13507, Pa_Term) <br><br>
+
- Transformation (<a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-Term.pdf">Pa_Term</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-
 +
 
 +
USP_Pa-I13507.pdf">Pa_I13507</a>) <br><br>
  
 
<br>
 
<br>
Line 696: Line 720:
 
     <td > 09/06
 
     <td > 09/06
 
<br>       
 
<br>       
−
- Inoculum (Pa_I13507,Pl_I13507,Pa_Term)<br><br>
+
- Inoculum (<a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>,<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-
 +
 
 +
USP_Pl-I13507.pdf">Pl_I13507</a>,<a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-Term.pdf">Pa_Term</a>)<br><br>
 
<br>
 
<br>
 
     </td>
 
     </td>
Line 707: Line 733:
 
     <td valign=top align=left> 10/08
 
     <td valign=top align=left> 10/08
 
<br>       
 
<br>       
−
- miniprep (Pa_I13507, Pl_I13507, Pa_Term)<br><br>
+
- miniprep (<a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>,<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-
−
- NotI digestion (Pa_I13507, Pa_Term)<br><br>
+
 
−
- Agarose gel (Pa_I13507, Pl_I13507, Pa_Term, Pl_RBS) - Positive results for Pa_I13507 and Pa_Term <br><br>
+
USP_Pl-I13507.pdf">Pl_I13507</a>,<a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-Term.pdf">Pa_Term</a>)<br><br>
 +
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>,<a href="https://static.igem.org/mediawiki/2015/4/40/Team-
 +
 
 +
Brasil-USP_Pa-Term.pdf">Pa_Term</a>)<br><br>
 +
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>,<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-
 +
 
 +
USP_Pl-I13507.pdf">Pl_I13507</a>,<a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-Term.pdf">Pa_Term</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/c/c6/Team-Brasil-USP_Pl_RBS.pdf">Pl_RBS</a>) - Positive results for Pa_I13507 and Pa_Term <br><br>
 
- Pre-inoculum of interlab study devices, BBa_K876004, and controls<br><br>
 
- Pre-inoculum of interlab study devices, BBa_K876004, and controls<br><br>
 
<br>
 
<br>
Line 715: Line 749:
 
     <td valign=top align=left> 11/08
 
     <td valign=top align=left> 11/08
 
<br>       
 
<br>       
−
- Vector exchange digestion (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term)<br><br>
+
- Vector exchange digestion (<a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>, <a
−
- Ligation with pSB1C3 (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term)<br><br>
+
 
−
- EXSP digestion (Pl_RBS, GFP_I13507, Pt_Term)<br><br>
+
href="https://static.igem.org/mediawiki/2015/4/4e/Team-Brasil-USP_GFP-I13507.pdf">GFP_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/8/8c/Team-Brasil-USP_GFP-
−
- ligation (Pl_I13507, Pl_Term)<br><br>
+
 
−
- Transformation OmpA<br><br>
+
P0440.pdf">GFP_P0440</a>, <a href="https://static.igem.org/mediawiki/2015/f/f1/Team-Brasil-USP_Tet-term.pdf">Tet_Term</a>)<br><br>
 +
- Ligation with pSB1C3 (<a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/4/4e/Team-
 +
 
 +
Brasil-USP_GFP-I13507.pdf">GFP_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/8/8c/Team-Brasil-USP_GFP-P0440.pdf">GFP_P0440</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/f/f1/Team-Brasil-USP_Tet-term.pdf">Tet_Term</a>)<br><br>
 +
- EXSP digestion (<a href="https://static.igem.org/mediawiki/2015/c/c6/Team-Brasil-USP_Pl_RBS.pdf">Pl_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/4/4e/Team-Brasil-
 +
 
 +
USP_GFP-I13507.pdf">GFP_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-USP_Pt-E0840.pdf">Pt_Term</a>)<br><br>
 +
- ligation (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-
 +
 
 +
USP_Pl-E0840.pdf">Pl_Term</a>)<br><br>
 +
- Transformation <a href="http://parts.igem.org/Part:BBa_K103006">OmpA</a><br><br>
 
- Sample preparation of Interlab Study devices and controls (plate reader)<br><br>
 
- Sample preparation of Interlab Study devices and controls (plate reader)<br><br>
 
- End-point measurement of Interlab Study devices and controls <br><br>
 
- End-point measurement of Interlab Study devices and controls <br><br>
−
- Sample preparation of BBa_K876004 part (plate reader)<br><br>
+
- Sample preparation of <a href="http://parts.igem.org/Part:BBa_K876004">BBa_K876004</a> part (plate reader)<br><br>
−
- Kinetic measurement of BBa_K876004 (14 hours).<br><br>
+
- Kinetic measurement of <a href="http://parts.igem.org/Part:BBa_K876004">BBa_K876004</a> (14 hours).<br><br>
 
<br>  
 
<br>  
 
     </td>
 
     </td>
Line 729: Line 775:
 
     <td valign=top align=left> 12/08
 
     <td valign=top align=left> 12/08
 
<br>
 
<br>
−
- Transformation (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term)<br><br>
+
- Transformation (<a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/4/4e/Team-Brasil-
−
- Inoculum OmpA<br><br>
+
 
−
- Pre-inoculum of interlab study devices, BBa_K876004, and controls      <br><br>
+
USP_GFP-I13507.pdf">GFP_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/8/8c/Team-Brasil-USP_GFP-P0440.pdf">GFP_P0440</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/f/f1/Team-Brasil-USP_Tet-term.pdf">Tet_Term</a>)<br><br>
 +
- Inoculum <a href="http://parts.igem.org/Part:BBa_K103006">OmpA</a><br><br>
 +
- Pre-inoculum of interlab study devices, <a href="http://parts.igem.org/Part:BBa_K876004">BBa_K876004</a>, and controls      <br><br>
 
<br>
 
<br>
 
        
 
        
 
     <td valign=top align=left> 13/08
 
     <td valign=top align=left> 13/08
 
<br>       
 
<br>       
−
- Inoculum (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term)<br><br>
+
- Inoculum (<a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/4/4e/Team-Brasil-USP_GFP-
−
- miniprep OmpA<br><br>
+
 
−
- EXSP digestion (B0034, OmpA)<br><br>
+
I13507.pdf">GFP_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/8/8c/Team-Brasil-USP_GFP-P0440.pdf">GFP_P0440</a>, <a
−
- agarose gel (B0034, OmpA)<br><br>
+
 
−
- Ligation RBS_OmpA (RBS + OmpA)<br><br>
+
href="https://static.igem.org/mediawiki/2015/f/f1/Team-Brasil-USP_Tet-term.pdf">Tet_Term</a>)<br><br>
 +
- miniprep <a href="http://parts.igem.org/Part:BBa_K103006">OmpA</a><br><br>
 +
- EXSP digestion (<a href="http://parts.igem.org/Part:BBa_B0034">B0034</a>, <a href="http://parts.igem.org/Part:BBa_K103006">OmpA</a>)<br><br>
 +
- agarose gel (<a href="http://parts.igem.org/Part:BBa_B0034">B0034</a>, <a href="http://parts.igem.org/Part:BBa_K103006">OmpA</a>)<br><br>
 +
- Ligation <a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a> (RBS + OmpA)<br><br>
 
- Sample preparation of Interlab Study devices and controls (flow cytometry)<br><br>
 
- Sample preparation of Interlab Study devices and controls (flow cytometry)<br><br>
 
- Measurement of Interlab Study devices and controls <br><br>
 
- Measurement of Interlab Study devices and controls <br><br>
−
- Sample preparation of BBa_K876004 part  (flow cytometry)<br><br>
+
- Sample preparation of <a href="http://parts.igem.org/Part:BBa_K876004">BBa_K876004</a> part  (flow cytometry)<br><br>
−
- Flow cytometry measurement of BBa_K876004 (14 hours).<br><br>
+
- Flow cytometry measurement of <a href="http://parts.igem.org/Part:BBa_K876004">BBa_K876004</a> (14 hours).<br><br>
  
 
<br>
 
<br>
Line 752: Line 806:
  
 
<br>       
 
<br>       
−
- miniprep (Pr_RBS, GFP_P0440(C3), Pl_I13507) - Positive results for Pl_I13507<br><br>  
+
- miniprep (<a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/8/8c/Team-Brasil-USP_GFP-
−
- and GFP_P0440<br><br>
+
 
−
- Transformation RBS_OmpA<br><br>
+
P0440.pdf">GFP_P0440</a>(C3), <a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>) - Positive results for Pl_I13507<br><br>  
 +
- and <a href="https://static.igem.org/mediawiki/2015/8/8c/Team-Brasil-USP_GFP-P0440.pdf">GFP_P0440</a><br><br>
 +
- Transformation <a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a><br><br>
 
<br>
 
<br>
 
      
 
      
Line 760: Line 816:
 
     <td valign=top align=left> 15/08
 
     <td valign=top align=left> 15/08
 
<br>       
 
<br>       
−
- Inoculum (RBS_OmpA, GFP_I13507(C3)<br><br>
+
- Inoculum (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a>, <a href="https://static.igem.org/mediawiki/2015/4/4e/Team-Brasil-
 +
 
 +
USP_GFP-I13507.pdf">GFP_I13507</a>(C3)<br><br>
 
<br>
 
<br>
  
Line 769: Line 827:
 
        
 
        
 
<br>
 
<br>
−
- miniprep (RBS_OmpA, GFP_I13507(C3))<br><br>
+
- miniprep (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a>, <a href="https://static.igem.org/mediawiki/2015/4/4e/Team-Brasil-
−
- NotI digestion (RBS_OmpA, GFP_I13507(C3))<br><br>
+
 
−
- Agarose gel (RBS_OmpA, GFP_I13507(C3))<br><br>
+
USP_GFP-I13507.pdf">GFP_I13507</a>(C3))<br><br>
−
- Ligation Pl_Term(Pl_RBS + Pl_Term)<br><br>
+
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a>, <a href="https://static.igem.org/mediawiki/2015/4/4e/Team-
 +
 
 +
Brasil-USP_GFP-I13507.pdf">GFP_I13507</a>(C3))<br><br>
 +
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a>, <a href="https://static.igem.org/mediawiki/2015/4/4e/Team-Brasil-
 +
 
 +
USP_GFP-I13507.pdf">GFP_I13507</a>(C3))<br><br>
 +
- Ligation <a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_Term</a>(Pl_RBS + Pl_Term)<br><br>
 
<br>
 
<br>
  
Line 781: Line 845:
 
     <td valign=top align=left> 17/08
 
     <td valign=top align=left> 17/08
 
<br>       
 
<br>       
−
- Inoculum (RBS_OmpA)<br><br>
+
- Inoculum (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a>)<br><br>
−
- Transformation (Pl_Term)<br><br>
+
- Transformation (<a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_Term</a>)<br><br>
 
<br>       
 
<br>       
  
Line 790: Line 854:
 
<br>       
 
<br>       
 
- IDT primers and gBlocks arrived<br><br>
 
- IDT primers and gBlocks arrived<br><br>
−
- miniprep (RBS_OmpA)<br><br>
+
- miniprep (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a>)<br><br>
−
- NotI digestion (RBS_OmpA)<br><br>
+
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a>)<br><br>
−
- Agarose gel (RBS_OmpA) - Postive<br><br>
+
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">RBS_OmpA</a>) - Postive<br><br>
−
- Vector exchange digestion (RBS_hokD, Pr_RBS)<br><br>
+
- Vector exchange digestion (<a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-USP_RBS-hokD.pdf">RBS_hokD</a>, <a
−
- Ligation RBS_hokD(C3), Pr_RBS(C3)<br><br>
+
 
−
- Inoculum (Pl_Term)<br><br>
+
href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>)<br><br>
 +
- Ligation <a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-USP_RBS-hokD.pdf">RBS_hokD(C3)</a>, <a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-
 +
 
 +
USP_Pr-RBS.pdf">Pr_RBS</a>(C3)<br><br>
 +
- Inoculum (<a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_Term</a>)<br><br>
 
<br>   
 
<br>   
 
      
 
      
 
     <td valign=top align=left> 19/08
 
     <td valign=top align=left> 19/08
 
<br>
 
<br>
−
- Transformation (RBS_hokD(C3), Pr_RBS(C3))<br><br>
+
- Transformation (<a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-USP_RBS-hokD.pdf">RBS_hokD</a>(C3), <a href="https://static.igem.org/mediawiki/2015/3/31/Team-
−
- NotI digestion (Pl_Term)<br><br>
+
 
−
- Agarose gel (Pl_Term)- positive!!!<br><br>
+
Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>(C3))<br><br>
 +
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_Term</a>)<br><br>
 +
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_Term</a>)- positive!!!<br><br>
 
<br>
 
<br>
  
Line 810: Line 880:
 
<br>
 
<br>
 
- gBlocks and Primers resuspension and quantification<br><br>
 
- gBlocks and Primers resuspension and quantification<br><br>
−
- EP digestion (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term)<br><br>
+
- EP digestion (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-
−
- Ligation Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term with pBS1C3<br><br>
+
 
−
- Inoculum (RBS_hokD(C3), Pr_RBS(C3))<br><br>
+
Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_term</a>, <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-
 +
 
 +
Term.pdf">Pa_Term</a>, <a href="https://static.igem.org/mediawiki/2015/4/4d/Team-Brasil-USP_Pr-E0840.pdf">Pr_Term</a>)<br><br>
 +
- Ligation <a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-
 +
 
 +
USP_Pa-I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_term</a>, <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-
 +
 
 +
Term.pdf">Pa_Term</a>, <a href="https://static.igem.org/mediawiki/2015/4/4d/Team-Brasil-USP_Pr-E0840.pdf">Pr_Term</a> with pBS1C3<br><br>
 +
- Inoculum (<a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-USP_RBS-hokD.pdf">RBS_hokD</a>(C3), <a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-
 +
 
 +
USP_Pr-RBS.pdf">Pr_RBS</a>(C3))<br><br>
 
<br>
 
<br>
 
        
 
        
Line 825: Line 909:
 
- Digestion of Lcp PCR biobrick and pSB1C3 with EP<br><br>
 
- Digestion of Lcp PCR biobrick and pSB1C3 with EP<br><br>
 
- Digestion of Lcp PCR for pETSUMO and pETSUMO with restriction enzymes BamHI and HindIII<br><br>
 
- Digestion of Lcp PCR for pETSUMO and pETSUMO with restriction enzymes BamHI and HindIII<br><br>
−
- miniprep (RBS_hokD(C3), Pr_RBS(C3))<br><br>
+
- miniprep (<a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-USP_RBS-hokD.pdf">RBS_hokD</a>(C3), <a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-
−
- NotI digestion (RBS_hokD(C3), Pr_RBS(C3))<br><br>
+
 
−
- Agarose gel (RBS_hokD(C3), Pr_RBS(C3)) - All negative<br><br>
+
USP_Pr-RBS.pdf">Pr_RBS</a>(C3))<br><br>
−
- Transformation (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term)<br><br>
+
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-USP_RBS-hokD.pdf">RBS_hokD</a>(C3), <a href="https://static.igem.org/mediawiki/2015/3/31/Team-
−
- Pre-inoculum (Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls)<br><br>
+
 
 +
Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>(C3))<br><br>
 +
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/d/d5/Team-Brasil-USP_RBS-hokD.pdf">RBS_hokD</a>(C3), <a href="https://static.igem.org/mediawiki/2015/3/31/Team-
 +
 
 +
Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>(C3)) - All negative<br><br>
 +
- Transformation (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-
 +
 
 +
Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_term</a>, <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-
 +
 
 +
Term.pdf">Pa_Term</a>, <a href="https://static.igem.org/mediawiki/2015/4/4d/Team-Brasil-USP_Pr-E0840.pdf">Pr_Term</a>)<br><br>
 +
- Pre-inoculum (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-
 +
 
 +
Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, interlab study devices and  
 +
 
 +
controls)<br><br>
 
<br>
 
<br>
 
     </td>
 
     </td>
Line 835: Line 935:
 
     <td valign=top align=left> 22/08
 
     <td valign=top align=left> 22/08
 
<br>       
 
<br>       
−
- Agarose gel purification for Lcp ligations<br><br>
+
- Agarose gel purification for <a href="http://parts.igem.org/Part:BBa_K1819001">Lcp</a> ligations<br><br>
−
- Transformation of Lcp for biobrick and Lcp for pETSUMO using insert and vector purified<br><br>
+
- Transformation of <a href="http://parts.igem.org/Part:BBa_K1819001">Lcp</a> for biobrick and Lcp for pETSUMO using insert and vector purified<br><br>
 
- Inoculum of 6 roxA mutant colonies<br><br>
 
- Inoculum of 6 roxA mutant colonies<br><br>
−
- Inoculum (Pl_I13507(C3))<br><br>
+
- Inoculum (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>(C3))<br><br>
−
- Sample preparation (Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls) (plate reader) - five 96-well plates<br><br>
+
- Sample preparation (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a
−
- Measurement of Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls (12 hours - plate reader)<br><br>
+
 
−
- Pre-inoculum (Pl_Term, Pa_Term, Pr_Term, BBa_K876004 and controls) (codes))<br><br>
+
href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-
 +
 
 +
I13507.pdf">Pr_I13507</a>, interlab study devices and controls) (plate reader) - five 96-well plates<br><br>
 +
- Measurement of <a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-
 +
 
 +
Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, interlab study devices and  
 +
 
 +
controls (12 hours - plate reader)<br><br>
 +
- Pre-inoculum (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-
 +
 
 +
Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, <a
 +
 
 +
href="http://parts.igem.org/Part:BBa_K876004">BBa_K876004</a> and controls) (codes))<br><br>
 
<br>
 
<br>
 
      
 
      
 
  <td valign=top align=left> 23/08
 
  <td valign=top align=left> 23/08
 
<br>       
 
<br>       
−
- gBlock reactions and transformations (linker-GFP, 6XHis, signalTAT)<br><br>
+
- gBlock reactions and transformations (<a href="http://parts.igem.org/Part:BBa_K1819000">linker-GFP</a>, 6XHis, <a
 +
 
 +
href="http://parts.igem.org/Part:BBa_K1819002">signalTAT</a>)<br><br>
 
- Miniprep roxA mutants<br><br>
 
- Miniprep roxA mutants<br><br>
 
- Digestion of EcoRI and HindIII of roxA mutants and plasmid non-mutated (control)<br><br>
 
- Digestion of EcoRI and HindIII of roxA mutants and plasmid non-mutated (control)<br><br>
 
- Agarose gel of the restriction analysis<br><br>
 
- Agarose gel of the restriction analysis<br><br>
−
- Sample preparation (Pl_Term, Pa_Term, Pr_Term, BBa_K876004 and controls) (plate reader) - five 96-well plates<br><br>
+
- Sample preparation (<a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_term</a>, <a href="https://static.igem.org/mediawiki/2015/4/40/Team-
−
- Measurement of Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507 and BBa_K876004 (12 hours - plate reader)<br><br>
+
 
−
- miniprep (Pl_I13507(C3))<br><br>
+
Brasil-USP_Pa-Term.pdf">Pa_Term</a>, <a href="https://static.igem.org/mediawiki/2015/4/4d/Team-Brasil-USP_Pr-E0840.pdf">Pr_Term</a>, <a
−
- NotI digestion (Pl_I13507(C3))<br><br>
+
 
−
- Agarose gel (Pl_I13507(C3)) - posite results<br><br>
+
href="http://parts.igem.org/Part:BBa_K876004">BBa_K876004</a> and controls) (plate reader) - five 96-well plates<br><br>
 +
- Measurement of <a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_term</a>, <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-
 +
 
 +
USP_Pa-Term.pdf">Pa_Term</a>, <a href="https://static.igem.org/mediawiki/2015/4/4d/Team-Brasil-USP_Pr-E0840.pdf">Pr_Term</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-
 +
 
 +
I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a> and <a
 +
 
 +
href="http://parts.igem.org/Part:BBa_K876004">BBa_K876004</a> (12 hours - plate reader)<br><br>
 +
- miniprep (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>(C3))<br><br>
 +
- NotI digestion (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>(C3))<br><br>
 +
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>(C3)) - posite results<br><br>
  
 
<br>
 
<br>
Line 864: Line 990:
 
     <td valign=top align=left> 24/08
 
     <td valign=top align=left> 24/08
 
<br>       
 
<br>       
−
- Inoculum (linker-GFP, 6XHis, signalTAT, Lcp(pETSUMO), Lcp(pSB1C3))<br><br>
+
- Inoculum (<a href="http://parts.igem.org/Part:BBa_K1819000">linker-GFP</a>, 6XHis, <a href="http://parts.igem.org/Part:BBa_K1819002">signalTAT</a>, Lcp(pETSUMO), <a
−
- miniprep (Pa_Term, Pl_Term)<br><br>
+
 
−
- EP digestion (Pa_Term, Pr_Term, Pl_Term, 2xHokD, B0015)<br><br>
+
href="http://parts.igem.org/Part:BBa_K1819001">Lcp</a>(pSB1C3))<br><br>
 +
- miniprep (<a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_term</a>, <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-
 +
 
 +
USP_Pa-Term.pdf">Pa_Term</a>)<br><br>
 +
- EP digestion (<a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_term</a>, <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-
 +
 
 +
USP_Pa-Term.pdf">Pa_Term</a>, <a href="https://static.igem.org/mediawiki/2015/4/4d/Team-Brasil-USP_Pr-E0840.pdf">Pr_Term</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/5/53/Team-Brasil-USP_2xrbs-hokD.pdf">2xHokD</a>, <a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>)<br><br>
 
- PCR of mutant roxA for biobrick ligation<br><br>
 
- PCR of mutant roxA for biobrick ligation<br><br>
 
<br>
 
<br>
Line 874: Line 1,008:
  
 
<br>       
 
<br>       
−
- miniprep (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3))<br><br>
+
- miniprep (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">OmpA_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-
−
- NotI and EXSP digestions ( (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3))<br><br>
+
 
−
- Agarose gel ((OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)) - Positive results for OmpA_RBS ,linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)<br><br>
+
USP_Pr-RBS.pdf">Pr_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, <a
−
- Ligation 6xHis_Term (linker6xHis +B0015), GFP_Term (linkerGFP + B0015), OmpA_Lcp (OmpA + Lcp(pSB1C3)), Tat_Lcp (signalTAT + Lcp(pSB1C3)<br><br>
+
 
−
- Ligation (Pa_Term(C3), Pl_Term(C3), Pr_Term(C3)<br><br>
+
href="http://parts.igem.org/Part:BBa_K1819000">linker-GFP</a>, <a href="http://parts.igem.org/Part:BBa_K1819002">signalTAT</a>, 6xHis, Lcp(pETSUMO), <a
 +
 
 +
href="http://parts.igem.org/Part:BBa_K1819001">Lcp</a>(pSB1C3))<br><br>
 +
- NotI and EXSP digestions ( (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">OmpA_RBS</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-USP_Pr-RBS.pdf">Pr_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-
 +
 
 +
I13507.pdf">Pr_I13507</a>, <a href="http://parts.igem.org/Part:BBa_K1819000">linker-GFP</a>, <a href="http://parts.igem.org/Part:BBa_K1819002">signalTAT</a>, 6xHis,  
 +
 
 +
Lcp(pETSUMO), <a href="http://parts.igem.org/Part:BBa_K1819001">Lcp</a>(pSB1C3))<br><br>
 +
- Agarose gel (<a href="https://static.igem.org/mediawiki/2015/2/24/Team-Brasil-USP_RBS-OmpA.pdf">OmpA_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/3/31/Team-Brasil-
 +
 
 +
USP_Pr-RBS.pdf">Pr_RBS</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, <a
 +
 
 +
href="http://parts.igem.org/Part:BBa_K1819000">linker-GFP</a>, <a href="http://parts.igem.org/Part:BBa_K1819002">signalTAT</a>, 6xHis, Lcp(pETSUMO), <a
 +
 
 +
href="http://parts.igem.org/Part:BBa_K1819001">Lcp</a>(pSB1C3)) - Positive results for OmpA_RBS ,linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)<br><br>
 +
- Ligation 6xHis_Term (linker6xHis +B0015), <a href="https://static.igem.org/mediawiki/2015/7/77/Team-Brasil-USP_linker-Term.pdf">GFP_Term</a> (linkerGFP + B0015), <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a> (OmpA + Lcp(pSB1C3)), <a href="https://static.igem.org/mediawiki/2015/7/74/Team-
 +
 
 +
Brasil-USP_TAT-Lcp.pdf">Tat_Lcp</a> (signalTAT + Lcp(pSB1C3)<br><br>
 +
- Ligation (<a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-Term.pdf">Pa_Term</a>(C3), <a href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-
 +
 
 +
USP_Pl-E0840.pdf">Pl_Term</a>(C3), <a href="https://static.igem.org/mediawiki/2015/4/4d/Team-Brasil-USP_Pr-E0840.pdf">Pr_Term</a>(C3)<br><br>
 
- Agarose gel (mutant RoxA)<br><br>
 
- Agarose gel (mutant RoxA)<br><br>
 
<br>
 
<br>
Line 885: Line 1,043:
 
     <td valign=top align=left> 26/08
 
     <td valign=top align=left> 26/08
 
<br>       
 
<br>       
−
- Transformation 6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp<br><br>
+
- Transformation6xHis_Term (linker6xHis +B0015), <a href="https://static.igem.org/mediawiki/2015/7/77/Team-Brasil-USP_linker-Term.pdf">GFP_Term</a> (linkerGFP + B0015), <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a> (OmpA + Lcp(pSB1C3)), <a href="https://static.igem.org/mediawiki/2015/7/74/Team-
 +
 
 +
Brasil-USP_TAT-Lcp.pdf">Tat_Lcp</a> (signalTAT + Lcp(pSB1C3)<br><br>
 
<br>
 
<br>
 
      
 
      
Line 892: Line 1,054:
 
     <td valign=top align=left> 27/08
 
     <td valign=top align=left> 27/08
 
<br>       
 
<br>       
−
- Inoculum (6xHis_Term, GFP_Term, mpA_Lcp, Tat_Lcp)<br><br>
+
- Inoculum (6xHis_Term (linker6xHis +B0015), <a href="https://static.igem.org/mediawiki/2015/7/77/Team-Brasil-USP_linker-Term.pdf">GFP_Term</a> (linkerGFP + B0015), <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a> (OmpA + Lcp(pSB1C3)), <a href="https://static.igem.org/mediawiki/2015/7/74/Team-
 +
 
 +
Brasil-USP_TAT-Lcp.pdf">Tat_Lcp</a> (signalTAT + Lcp(pSB1C3))<br><br>
 
- Transformation (2xhokD_Term)<br><br>
 
- Transformation (2xhokD_Term)<br><br>
 
- PCR of mutant roxA for biobrick ligation<br><br>
 
- PCR of mutant roxA for biobrick ligation<br><br>
Line 903: Line 1,069:
 
- roxA PCR product purification<br><br>
 
- roxA PCR product purification<br><br>
 
- roxA PCR product digestion with EP<br><br>
 
- roxA PCR product digestion with EP<br><br>
−
- miniprep (6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp)<br><br>
+
- miniprep (6xHis_Term (linker6xHis +B0015), <a href="https://static.igem.org/mediawiki/2015/7/77/Team-Brasil-USP_linker-Term.pdf">GFP_Term</a> (linkerGFP + B0015), <a
−
- NotI and EXSP digestions (6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp) - Only 6xHis_Term was negative<br><br>
+
 
 +
href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a> (OmpA + Lcp(pSB1C3)), <a href="https://static.igem.org/mediawiki/2015/7/74/Team-
 +
 
 +
Brasil-USP_TAT-Lcp.pdf">Tat_Lcp</a> (signalTAT + Lcp(pSB1C3))<br><br>
 +
- NotI and EXSP digestions (6xHis_Term (linker6xHis +B0015), <a href="https://static.igem.org/mediawiki/2015/7/77/Team-Brasil-USP_linker-Term.pdf">GFP_Term</a> (linkerGFP +
 +
 
 +
B0015), <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a> (OmpA + Lcp(pSB1C3)), <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">Tat_Lcp</a> (signalTAT + Lcp(pSB1C3)) - Only 6xHis_Term was negative<br><br>
 
- ligation Lcp_Term I (Tat_Lcp + GFP_Term), Lcp_Term II (OmpA_Lcp + GFP_Term) <br><br>
 
- ligation Lcp_Term I (Tat_Lcp + GFP_Term), Lcp_Term II (OmpA_Lcp + GFP_Term) <br><br>
−
- Pre-inoculum (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507)<br><br>
+
- Pre-inoculum (<a href="https://static.igem.org/mediawiki/2015/4/41/Team-Brasil-USP_Pl-I13507.pdf">Pl_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/0/04/Team-
 +
 
 +
Brasil-USP_Pa-I13507.pdf">Pa_I13507</a>, <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>, <a
 +
 
 +
href="https://static.igem.org/mediawiki/2015/d/d8/Team-Brasil-USP_Pl-E0840.pdf">Pl_term</a>, <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-
 +
 
 +
Term.pdf">Pa_Term</a>, <a href="https://static.igem.org/mediawiki/2015/4/4d/Team-Brasil-USP_Pr-E0840.pdf">Pr_Term</a>)<br><br>
  
 
<br>
 
<br>
 
     </td>
 
     </td>
 +
         
 +
 
            
 
            
 
           <td valign=top align=left> 29/08
 
           <td valign=top align=left> 29/08

Revision as of 07:48, 18 September 2015

Notebook

Calendar

Table of contents

June

Monday Tuesday Wednesday Thursday Friday Saturday Sunday
01/06 02/06
- iGEM kit arrived! 		03/06 04/06 05/06 06/06 07/06
08/06 09/06
- Transformation:
- B0034
- K1497008
- R0010
- K914003
- R0080
- C0040
- B0015
- R0040
- E0840
- E0040
- P0440
- I13507
 10/06 11/06
- Inoculum
- B0034
- K1497008
- R0010
- K914003
- R0080
- C0040
- B0015
- R0040
- E0840
- E0040
- P0440
- I13507
 12/06
- Miniprep and Digestion EXSP
- B0034
- K1497008
- R0010
- K914003
- R0080
- C0040
- B0015
- R0040
- E0840
- E0040
- P0440
- I13507
 13/06
- Agarose gel (Problems with the buffer! Way too concentrated ><)
- Digestion EXSP again (C0040, B0015, R0040, E0840, E0040, P0440, I13507) [We didn’t have B0034 anymore, so we didn’t prepare the reaction for K1497008, R0010, K914003, R0080] 		14/06
- Agarose gel(C0040, B0015, R0040, E0840, E0040, P0440, I13507)
- Gel purification (C0040, B0015, E0040, P0440)
- Interlab transformations (J23101, J23106, J23117, I13504)
- Ligation Tet_term (C0040 + B0015)
- Ligation GFP_P0440 (E0040 + P0440)
15/06
- Transformation Tet_term (C0040 + B0015)
- Transformation GFP_P0440 (E0040 + P0440)
- Interlab inoculum
- Inoculum (B0034) 		16/06
- Inoculum (Tet_term, GFP_P0440)
- Interlab minipreps
- Miniprep (B0034)
- Digestion ESXP (B0034, K1497008, R0010, K914003, R0040, E0840) 		17/06
- Miniprep (Tet_term, GFP_P0440)
- Agarose gel (B0034, K1497008, R0010, K914003, R0040, E0840)
- Gel purification (B0034, K1497008, R0010, K914003, R0040, E0840)
- Ligation RBS_hokD (B0034 + K1497008)
- Ligation Pl_RBS (R0010 + B0034)
- Ligation Pt_E0840 (R0040 + E0840)
- Ligation Pt_I13507 (R0040 + I13507) 		18/06
- Transformation (RBS_hokD, Pl_RBS, Pt_E0840, Pt_I13507) 		19/06
- No colonies! Wrong antibiotic!!!!
- New transformation (RBS_hokD, Pl_RBS, Pt_E0840, Pt_I13507) 		20/06
- Inoculum (Pl_RBS, Pt_E0840, Pt_I13507)
- RBS_hokD didn’t grow 		21/06
- Miniprep (Pl_RBS, Pt_E0840, Pt_I13507)
- Digestion NotI (Tet_term, GFP_P0440, Pl_RBS, Pt_E0840, Pt_I13507)
22/06
- Confirmation, agarose gel (Tet_term, GFP_P0440, Pl_RBS, Pt_E0840, Pt_I13507)
- Confirmed (Tet_term, Pl_RBS, Pt_E0840)
- Ligation (GFP_P0440, Pt_I13507, RBS_hokD) 		23/06
- Transformation (GFP_P0440, Pt_I13507, RBS_hokD) 		24/06
- Inoculum [GFP_P0440 (3 colonies), RBS_hokD (3 colonies), I13507, R0080, K914003] 		25/06
- Miniprep [GFP_P0440 (3 colonies), RBS_hokD (3 colonies), I13507, R0080, K914003] 		26/06 27/06
- EXSP digestion (K914003)
- Confirmation digestion (RBS_hokD) 		28/06
- Agarose gel (K914003)
- Agarose gel (RBS_hokD) – positive result \o/
- Gel purification (K914003)
- Ligation Pr_RBS (K91403 + B0034) 22ºC for 4h, 16ºC for 10h, 4º overnight
- EXSP digestion RBS_hokD
- Interlab digestion (from minipreps of June 15th)
- Promoters J23101,J23106 and J23117 - SP enzymes
29/06
- Agarose gel RBS_hokD
- Ligation (Pr_RBS, Pa_RBS) 		30/06
- Inoculum RBS_hokD
- EXSP digestion RBS_hokD

July

Monday Tuesday Wednesday Thursday Friday Saturday Sunday
01/07
- Agarose gel (RBS_hokD)

- Gel purification (RBS_hokD)

- Ligation 2X_RBS_hokD (RBS_hokD + RBS_hokD)

- EXSP Digestion (R0040, Pl_RBS, GFP_P0440)

- Agarose gel (R0040)

- Gel purification (R0040)

- Ligation Pt_I13507(R0040 + I13507)

- Interlab gel purification and ligation


02/07
- Transformation Ptet_I13507

- Agarose gel (Ptet_I13507, GFP_P0440)

- Gel purification (Pl_RBS, GFP_P0440)

- Ligation Pl_P0440 (Pl_RBS + GFP_P0440)

- Interlab transformation (with wrong antibiotic ampicillin - it is possible to find in registry the same parts in another backbone:BBa_J61002)


 03/07
- Transformation (Pr_RBS, 2X_RBS_hokD)

- EXSP digestion (P0440, R0040)

- Transformation Pl_P0440 (Pl_RBS + GFP_P0440)

- Inoculum (Pt_I13507)


 04/07
- Agarose gel (P0440, R0040)

- Gel purification (P0440, R0040)

- Ligation tet_Ptet (P0440 + R0040)

- Inoculum (2X_RBS_hokD)

- Miniprep (Pt_I13507)


 05/07
- Inoculum (R0080, K91403)

- EXSP digestion (tet_Term, Pt_E0840)

- Miniprep (2X_RBS_hokD)
06/07
- NotI digestion (2X_RBS_hokD, Pr_RBS, Pa_RBS)

- Transformation (tet_Ptet)

- Agarose gel (2X_RBS_hokD) – positive results

- Agarose gel (tet_Term, Pt_E0840)

- EXSP digestion (Pt_E0840)

- Inoculum (Pl_P0440)

- NotI digestion (Pt_I13507) – positive results


07/07
- Agarose gel (Pr_RBS, Pa_RBS, Pt_E0480) – positive results for Pr_RBS and Pt_E0840)

- EXSP digestion (2X_RBS_hokD, B0015)

- Ligation Term_E0840 (Pt_E0840 + tet_Term)

- Inoculum (tet_Ptet)

- Transformation (Term_E0840)

- Miniprep (Pl_P0440)

- Not digestion (Pl_P0440) – positive results

- EXSP digestion (Pl_P0440, Pt_I13507)

- Inoculum (GFP_P0440)


 08/07
- Agarose gel (2X_RBS_hokD, B0015) – B0015 didn’t digest

- Gel purification (2X_RBS_hokD)

- EXSP digestion (B0015)

- Miniprep (tet_Ptet)

- NotI digestion (tet_Ptet)

- Agarose gel (tet_Ptet, B0015) – positive results

- Gel digestion (B0015)

- Ligation hokD_Term (2X_RBS_hokD + B0015)

- Ligation Pl_I13507 (Pl_P0440 + Pt_I13507)

- Miniprep (GFP_P0440)

- EXSP digestion (GFP_P0440)


 09/07
- Agarose gel and gel purification (GFP_P0440)

- Ligation GFP_I13507 (GFP_P0440 + Pt_I13507)

- Transformation (Pl_I13507)


 10/07
- Transformation (hokD_Term)

- Inoculum (Pl_I13507)

- Transformation (GFP_I13507)


 11/07
- Miniprep (Pl_I13507)


 12/07
13/07
- Inoculum (GFP_I13507)

- NotI digestion (Pl_I13507)

- Agarose gel (Pl_I13507) – negative results


 14/07
- Miniprep (GFP_I13507)

- NotI digestion (GFP_I13507) – negative results

- EXSP digestion (Pl_P0440, Pt_I13507)

- Inoculum (GFP_P0440, Pt_I13507)


 15/07
- Agarose gel (Pl_P0440, Pt_I13507)

- Gel purification (Pl_P0440, Pt_I13507)

- Ligation Pl_I13507 (Pl_P0440 + Pt_I13507)

- Miniprep (GFP_P0440, Pt_I13507)

- EXSP digestion (GFP_P0440, Pt_I13507)


 16/07
- Agarose gel and gel purification (GFP_P0440, Pt_I13507)

- Ligation GFP_I13507 (GFP_P0440 + Pt_I13507)

- Transformation (Pl_I13507)


 17/07
- Transformation (GFP_I13507)

- Inoculum (Pt_Term, Pl_I13507)


 18/07
- Miniprep (Pt_Term, Pl_I13507)

- NotI digestion (Pt_Term, Pl_I13507)

- Agarose gel (Pt_Term, Pl_I13507) – positive results for Pt_Term


 19/07
20/07
  • iGEM kit arrived!
    		•  21/07
    - Inoculum (GFP_I13507, Pl_I13507)

    - New interlab transformations


    		 22/07
    - Miniprep (GFP_I13507, Pl_I13507)

    - NotI digestion (GFP_I13507, Pl_I13507)

    - EXSP digestion (Pr_RBS, R0080)

    - Agarose gel (GFP_I13507, Pl_I13507, Pr_RBS)

    - Gel purification (Pr_RBS)

    - Inoculum (GFP_I13507, Pl_I13507)

    - Interlab inocula


    		 23/07
    - Agarose gel (R0080)

    - Miniprep (GFP_I13507, Pl_I13507)

    - NotI digestion (GFP_I13507, Pl_I13507)

    - Agarose gel (GFP_I13507, Pl_I13507) – Positive results for GFP_I13507

    - EXSP digestion (GFP_I13507)

    - Interlab miniprep and restriction analysis


    		 24/07
    - Miniprep (B0015, Pl_P0440)

    - EXSP digestion (2X_RBS_hokD, B0015)

    - Transformation (Pa_RBS)

    - Agarose gel of restriction analysis: devices confirmed


    		 25/07
    - agarose gel (2X_RBS_hokD, B0015) - B0015 did not digest


    		 26/07
    - Agarose gel and gel purification (GFP_I13507)

    - Ligation Pr_I13507 (Pr_RBS + GFP_I13507)


    27/07
    - EXSP digestion (Pl_RBS, Pt_Term)

    - Transformation (Pr_I13507, Pa_RBS)


    		 28/07
    - Agarose gel (Pl_RBS, Pt_Term)

    ligation Pr_I13507(Pr_RBS + Pt_Term)


    		 29/07
    - EXSP digestion (B0015, 2X_RBS_hokD)

    - agarose gel (B0015, 2X_RBS_hokD) - 2X_RBS_hokD didn’t digest

    - EXSP digestion overnight (2X_RBS_hokD)

    - transformation (Pr_Term, Pl_RBS)


    		 30/07
    - inoculum (Pl_RBS, Pa_RBS)

    - agarose gel (B0015, 2X_RBS_hokD) - Digestion worked!

    - ligation 2X_hokD_Term (B0015 + 2X_RBS_hokD) P0440_hokD(P0440_term + 2X_RBS_hokD)


    		 31/07

    August

    Monday Tuesday Wednesday Thursday Friday Saturday Sunday
    01/08 02/08
    03/08
    - EXSP digestion (R0010, R0080, B0034, Pt_I13507)

    - NotI digestion (Pl_I13507)

    - Ligation Pa_RBS (R0080 + B0034) and Pl_RBS (R0010 + B0034)


    		 04/08
    - NotI digestion (Pr_I13507)

    - Agarose gel (Pr_I13507) - Positive results!!

    - EXSP digestion (Pt_I13507)

    - Transformation (Pl_RBS and Pa_RBS)


    		 05/08
    - agarose gel (Pl_I13507, Pt_I13507)- negative results for Pl_I13507 :(

    - Agarose gel and gel purification (Pt_I13507)

    - ligation Pl_I13507 (Pl_P0440 + Pt_I13507)

    - inoculum (Pl_RBS, Pa_RBS)


    		 06/08
    - Transformation (Pl_I13507, Pr_I13507)

    - miniprep (Pl_RBS, Pa_RBS)

    - NotI digestion (Pl_RBS, Pa_RBS)


    		 07/08
    - Agarose gel (Pl_RBS, Pa_RBS) - Only Pa_RBS was positive

    - EXSP digestion (Pa_RBS, Pt_Term)

    - Ligation Pa_Term(Pa_RBS + Pt_Term), Pa_I13507 (Pa_RBS + GFP_I13507)


    		 08/08
    - Ligation (2X_RBS_hokD + Term)

    - Transformation (Pa_Term, Pa_I13507)


    		09/06
    - Inoculum (Pa_I13507,Pl_I13507,Pa_Term)
    10/08
    - miniprep (Pa_I13507,Pl_I13507,Pa_Term)

    - NotI digestion (Pa_I13507,Pa_Term)

    - Agarose gel (Pa_I13507,Pl_I13507,Pa_Term, Pl_RBS) - Positive results for Pa_I13507 and Pa_Term

    - Pre-inoculum of interlab study devices, BBa_K876004, and controls


    		 11/08
    - Vector exchange digestion (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term)

    - Ligation with pSB1C3 (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term)

    - EXSP digestion (Pl_RBS, GFP_I13507, Pt_Term)

    - ligation (Pl_I13507, Pl_Term)

    - Transformation OmpA

    - Sample preparation of Interlab Study devices and controls (plate reader)

    - End-point measurement of Interlab Study devices and controls

    - Sample preparation of BBa_K876004 part (plate reader)

    - Kinetic measurement of BBa_K876004 (14 hours).


    		 12/08
    - Transformation (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term)

    - Inoculum OmpA

    - Pre-inoculum of interlab study devices, BBa_K876004, and controls


    		13/08
    - Inoculum (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term)

    - miniprep OmpA

    - EXSP digestion (B0034, OmpA)

    - agarose gel (B0034, OmpA)

    - Ligation RBS_OmpA (RBS + OmpA)

    - Sample preparation of Interlab Study devices and controls (flow cytometry)

    - Measurement of Interlab Study devices and controls

    - Sample preparation of BBa_K876004 part (flow cytometry)

    - Flow cytometry measurement of BBa_K876004 (14 hours).


    		 14/08
    - miniprep (Pr_RBS, GFP_P0440(C3), Pl_I13507) - Positive results for Pl_I13507

    - and GFP_P0440

    - Transformation RBS_OmpA


    		 15/08
    - Inoculum (RBS_OmpA, GFP_I13507(C3)


    		 16/08
    - miniprep (RBS_OmpA, GFP_I13507(C3))

    - NotI digestion (RBS_OmpA, GFP_I13507(C3))

    - Agarose gel (RBS_OmpA, GFP_I13507(C3))

    - Ligation Pl_Term(Pl_RBS + Pl_Term)


    17/08
    - Inoculum (RBS_OmpA)

    - Transformation (Pl_Term)


    		 18/08
    - IDT primers and gBlocks arrived

    - miniprep (RBS_OmpA)

    - NotI digestion (RBS_OmpA)

    - Agarose gel (RBS_OmpA) - Postive

    - Vector exchange digestion (RBS_hokD, Pr_RBS)

    - Ligation RBS_hokD(C3), Pr_RBS(C3)

    - Inoculum (Pl_Term)


    		 19/08
    - Transformation (RBS_hokD(C3), Pr_RBS(C3))

    - NotI digestion (Pl_Term)

    - Agarose gel (Pl_Term)- positive!!!


    		 20/08
    - gBlocks and Primers resuspension and quantification

    - EP digestion (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term)

    - Ligation Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term with pBS1C3

    - Inoculum (RBS_hokD(C3), Pr_RBS(C3))


    		 21/08
    - PCR for mutation of EcoRI roxA site

    - Agarose gel for amplification confirmation and purification of roxA PCR mutation

    - DpnI digestion of purificated PCR and DH5a transformation using the DNA digested

    - PCR for insert amplification of Lcp inserting Biobrick sites

    - PCR for insert amplification of Lcp inserting pETSUMO restriction sites (Lcp heterologous expression)

    - Agarose gel for amplification confirmation and purification of lcp PCRs

    - Digestion of Lcp PCR biobrick and pSB1C3 with EP

    - Digestion of Lcp PCR for pETSUMO and pETSUMO with restriction enzymes BamHI and HindIII

    - miniprep (RBS_hokD(C3), Pr_RBS(C3))

    - NotI digestion (RBS_hokD(C3), Pr_RBS(C3))

    - Agarose gel (RBS_hokD(C3), Pr_RBS(C3)) - All negative

    - Transformation (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term)

    - Pre-inoculum (Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls)


    		 22/08
    - Agarose gel purification for Lcp ligations

    - Transformation of Lcp for biobrick and Lcp for pETSUMO using insert and vector purified

    - Inoculum of 6 roxA mutant colonies

    - Inoculum (Pl_I13507(C3))

    - Sample preparation (Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls) (plate reader) - five 96-well plates

    - Measurement of Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls (12 hours - plate reader)

    - Pre-inoculum (Pl_I13507, Pa_I13507, Pr_I13507, BBa_K876004 and controls) (codes))


    		 23/08
    - gBlock reactions and transformations (linker-GFP, 6XHis, signalTAT)

    - Miniprep roxA mutants

    - Digestion of EcoRI and HindIII of roxA mutants and plasmid non-mutated (control)

    - Agarose gel of the restriction analysis

    - Sample preparation (Pl_term, Pa_Term, Pr_Term, BBa_K876004 and controls) (plate reader) - five 96-well plates

    - Measurement of Pl_term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507 and BBa_K876004 (12 hours - plate reader)

    - miniprep (Pl_I13507(C3))

    - NotI digestion (Pl_I13507(C3))

    - Agarose gel (Pl_I13507(C3)) - posite results


    24/08
    - Inoculum (linker-GFP, 6XHis, signalTAT, Lcp(pETSUMO), Lcp(pSB1C3))

    - miniprep (Pl_term, Pa_Term)

    - EP digestion (Pl_term, Pa_Term, Pr_Term, 2xHokD, B0015)

    - PCR of mutant roxA for biobrick ligation


    		 25/08
    - miniprep (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3))

    - NotI and EXSP digestions ( (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3))

    - Agarose gel (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)) - Positive results for OmpA_RBS ,linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)

    - Ligation 6xHis_Term (linker6xHis +B0015), GFP_Term (linkerGFP + B0015), OmpA_Lcp (OmpA + Lcp(pSB1C3)), Tat_Lcp (signalTAT + Lcp(pSB1C3)

    - Ligation (Pa_Term(C3), Pl_Term(C3), Pr_Term(C3)

    - Agarose gel (mutant RoxA)


    		 26/08
    - Transformation6xHis_Term (linker6xHis +B0015), GFP_Term (linkerGFP + B0015), OmpA_Lcp (OmpA + Lcp(pSB1C3)), Tat_Lcp (signalTAT + Lcp(pSB1C3)


    		 27/08
    - Inoculum (6xHis_Term (linker6xHis +B0015), GFP_Term (linkerGFP + B0015), OmpA_Lcp (OmpA + Lcp(pSB1C3)), Tat_Lcp (signalTAT + Lcp(pSB1C3))

    - Transformation (2xhokD_Term)

    - PCR of mutant roxA for biobrick ligation

    - Agarose gel of mutant roxA PCR product


    		28/08
    - roxA PCR product purification

    - roxA PCR product digestion with EP

    - miniprep (6xHis_Term (linker6xHis +B0015), GFP_Term (linkerGFP + B0015), OmpA_Lcp (OmpA + Lcp(pSB1C3)), Tat_Lcp (signalTAT + Lcp(pSB1C3))

    - NotI and EXSP digestions (6xHis_Term (linker6xHis +B0015), GFP_Term (linkerGFP + B0015), OmpA_Lcp (OmpA + Lcp(pSB1C3)), Tat_Lcp (signalTAT + Lcp(pSB1C3)) - Only 6xHis_Term was negative

    - ligation Lcp_Term I (Tat_Lcp + GFP_Term), Lcp_Term II (OmpA_Lcp + GFP_Term)

    - Pre-inoculum (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term)


    		 29/08
    - Agarose gel (roxA PCR product)

    - Ligation roxA(mutant) + pSB1C3

    - Sample preparation (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507) (plate reader) - seven 96-well plates

    - Measurement of test promoters (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507)


    		 30/08
    - Transformation (roxA)


    31/08
    - Inoculum (roxA)

    - Transformation (Lcp_Term I, Lcp_Term II)

    September

    Monday Tuesday Wednesday Thursday Friday Saturday Sunday
    01/09
    - miniprep (roxA)

    - Inoculum (Lcp_Term I)

    - Ligation (Lcp_Term II)


    		 02/09
    - miniprep (Lcp_Term I)

    - digestion (Lcp_Term I, roxA))

    - Transformation (Lcp_Term II)


    		 03/09
    - Transformation (Pr_RBS,2X_RBS_hokDLigation.)

    -Pre-inoculum (Pl_Term,Pa_Term,Pr_Term, Pl_I13507, Pa_I13507,Pr_I13507)


    		 04/09
    - Transformation (Pl_2xhokD)

    - Sample preparation of test promoters circuits (plate reader)

    - Measurement of test promoters circuits (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507)


    		 05/09
    - Inoculum (Pl_2xhokD)


    		 06/09
    - Transformation in Bl21(DE3) (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507)
    07/09
    - Inoculum (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507 in BL21(DE3))


    		 08/09
    - Miniprep (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507Pa_I13507, Pr_I13507, in BL21(DE3))

    - Digestion with SacI (OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA)

    - Digestion with SacI and XbaI (OmpA, signalTAT, Lcp, RoxA)

    - EXSP digestion (R0010, R0080, K914003)

    - NotI digestion (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507Pa_I13507, Pr_I13507, in BL21(DE3))

    - Agarose gel (OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA, OmpA, signalTAT, Lcp, RoxA, Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507) - Positive results for Pa_I13507 and Pr_I13507Pr_I13507)

    - Ligation OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA, Pl_I13507II (R0010 + GFP_I13507), Pl_TermII (R0010 + tet_Term), Pa_I13507II (R0080 + GFP_I13507), Pa_TermII (R0080 + tet_Term), Pr_I13507II (K914003 + GFP_I13507), Pr_TermII (K914003 + tet_Term),

    -Transformation (OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA, Pl_I13507II, Pa_I13507II, Pr_I13507II, Pl_TermII, Pa_TermII, Pr_TermII)


    		 09/09
    -Western blot to see if LCP was been expressed in pETSUMO

    - inoculum (Pl_I13507II, Pa_I13507II, Pl_TermII, Pa_TermII, Pr_TermII, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt)

    - Prepare plates for characterization (Pl_I13507 and Pr_I13507 in BL21(DE3)) 10th


    		 10/09
    - Miniprep (Pl_I13507II, Pa_I13507II, Pl_TermII, Pa_TermII, Pr_TermII, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt)

    -NotI digestion (Pl_I13507II, Pe_I13507II, Pl_TermII, Pa_TermII, Pr_TermII)

    - SacI digestion again (OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt)

    -Agarose gel (Pl_I13507II, Pe_I13507II, Pl_TermII, Pa_TermII, Pr_TermII, OmpA_Lcp,OmpA_roxA, TAT_LcpTAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt)

    -Ligation (OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, GFP_TermA (J23101 + linkerGFP_Term), GFP_TermB (J23117 + linkerGFP_Term), GFP_TermC (J23106 + linkerGFP_Term),

    -Transformation (OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, GFP_TermA, GFP_TermB, GFP_TermC


    		 11/09
    - Transformation (TAT_ lcp_RBS,TAT_ lcp,TAT_ roxA, OmpA_roxA_Pt)

    -Inoculum (GFP_TermA, GFP_TermB, GFP_TermC)


    		 12/09
    - Miniprep (GFP_TermA, GFP_TermB, GFP_TermC)

    - Not digestion (GFP_TermA, GFP_TermB, GFP_TermC)

    - Xbal and NotI digestion (RoxA, signalTAT and Lcp)

    -Agarose gel (GFP_TermA, GFP_TermB, GFP_TermC, RoxA, signalTAT and Lcp)

    -Ligation TAT_Lcp (signalTAT + Lcp), TAT_RoxA (signalTAT + roxA)


    		 13/09
    - Transformation (TAT_Lcp, TAT_roxA)
    14/09
    - EP digestion to change the vector (Pa_Term, signalTAT, OmpA, Lcp, RoxA)

    - SacI and EcoRI digestion (signalTat, OmpA)

    - SacI and PstI digestion (Lcp, RoxA)

    - Agarose gel ( Pa_Term, signalTAT, OmpA, Lcp, RoxA)

    -Ligation (pSB1A2 + RoxA, pSB1A2 + Lcp, pSB1A2 + OmpA and pSB1A2 + SignalTAT)

    -3A assembly (TAT_RoxA(pSB1A2 +signalTA+RoxA), TAT_Lcp(pSB1A2 +signalTA+Lcp),OmpA_RoxA (pSB1A2 +OmpA+RoxA),OmpA_Lcp( pSB1A2 +OmpA+Lcp))

    -Transformation (RoxA, Lcp, OmpA, SignalTAT, (OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA)

    - Inoculum (linkerGFP_J23117, linkerGFP_J23106)


    		 15/09
    - Ligation again (pSB1A2 + RoxA, pSB1A2 + Lcp, pSB1A2 + OmpA and pSB1A2 + SignalTAT)

    - 3A assembly again (TAT_RoxA(pSB1A2 +signalTA+RoxA),TAT_Lcp(pSB1A2 +signalTA+Lcp), OmpA_RoxA(pSB1A2 +OmpA+RoxA),OmpA_Lcp( pSB1A2 +OmpA+Lcp))

    - Sample preparation (linkerGFP_J23117, linkerGFP_J23106)

    - Measurements (linkerGFP_J23117, linkerGFP_J23106)


    		 16/09
    - Transformation (RoxA, Lcp, OmpA, SignalTAT, OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA)

    - Prepare the plates to submit the new parts

    - Lcp Expression using pETSUMO in Bl21(DE3)


    		 17/09
    - Inoculum (RoxA, Lcp, OmpA, SignalTAT, OmpA_Lcp,OmpA_roxA, TAT_LcpTAT_Lcp, TAT_roxA)

    - Cell lyses

    - Western blot (Lcp)


    		 18/09
    - Miniprep (RoxA, Lcp, OmpA, SignalTAT, OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA)

    - NotI digestion (RoxA, Lcp, OmpA, SignalTAT, OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA)

    - Agarose gel (RoxA, Lcp, OmpA, SignalTAT, OmpA_Lcp,OmpA_roxA, TAT_LcpTAT_Lcp, TAT_roxA)

    - Western blot (Lcp)

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