Difference between revisions of "Team:Brasil-USP/Notebook/Calendar"
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− | <td > 09/07 | + | <td valign=top align=left> 09/07 |
<br> | <br> | ||
- Agarose gel and gel purification (<a href="https://static.igem.org/mediawiki/2015/8/8c/Team-Brasil-USP_GFP-P0440.pdf">GFP_P0440</a>)<br><br> | - Agarose gel and gel purification (<a href="https://static.igem.org/mediawiki/2015/8/8c/Team-Brasil-USP_GFP-P0440.pdf">GFP_P0440</a>)<br><br> | ||
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</td> | </td> | ||
</table> | </table> | ||
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− | + | <a id="August"></a> | |
− | <h1>August</h1> | + | <h1>August</h1> |
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- 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> | - 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 (<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- | + | - 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> |
− | + | ||
− | RBS.pdf">Pa_RBS</a>)<br><br> | + | |
<br> | <br> | ||
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USP_Pt-E0840.pdf">Pt_Term</a>)<br><br> | 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 | + | - 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> | href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a> (Pa_RBS + GFP_I13507)<br><br> | ||
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</td> | </td> | ||
− | <td > 09/06 | + | <td valign=top align=left> 09/06 |
<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- | + | - 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> |
− | + | ||
− | 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> | ||
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<td valign=top align=left> 10/08 | <td valign=top align=left> 10/08 | ||
<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- | + | - 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-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> |
− | + | ||
− | 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- | - 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> | 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- | + | - 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> |
− | + | ||
− | 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> | ||
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<td valign=top align=left> 26/08 | <td valign=top align=left> 26/08 | ||
<br> | <br> | ||
− | - | + | - Transformation 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- | 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- | ||
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</td> | </td> | ||
− | + | <td> | |
+ | </td> | ||
+ | <td> | ||
+ | </td> | ||
+ | <td> | ||
+ | </td> | ||
+ | <td> | ||
+ | </td> | ||
+ | <td> | ||
+ | </td> | ||
+ | |||
</table> | </table> | ||
+ | |||
+ | |||
+ | <a id="September"></a> | ||
<h1>September</h1> | <h1>September</h1> | ||
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<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/2015/5/53/Team-Brasil-USP_2xrbs-hokD.pdf">2X_RBS_hokDLigation</a>.)<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/2015/5/53/Team-Brasil-USP_2xrbs-hokD.pdf">2X_RBS_hokDLigation</a>.)<br><br> |
− | -Pre-inoculum (<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>)<br><br> | + | -Pre-inoculum (<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>)<br><br> |
<br> | <br> | ||
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<td valign=top align=left> 08/09 | <td valign=top align=left> 08/09 | ||
<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>, <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> | + | - 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>, <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>, in BL21(DE3))<br><br> |
- Digestion with SacI (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> | - Digestion with SacI (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> | ||
- Digestion with SacI and XbaI (OmpA, signalTAT, Lcp, RoxA)<br><br> | - Digestion with SacI and XbaI (OmpA, signalTAT, Lcp, RoxA)<br><br> | ||
- EXSP digestion (R0010, R0080, K914003)<br><br> | - EXSP digestion (R0010, R0080, K914003)<br><br> | ||
− | - NotI 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/2015/4/41/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> | + | - NotI 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/2015/4/41/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>, in BL21(DE3))<br><br> |
− | - Agarose gel (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, OmpA, signalTAT, Lcp, RoxA, <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>) - Positive results for <a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a> and <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a> | + | - Agarose gel (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, OmpA, signalTAT, Lcp, RoxA, <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>) - Positive results for <a href="https://static.igem.org/mediawiki/2015/0/04/Team-Brasil-USP_Pa-I13507.pdf">Pa_I13507</a> and <a href="https://static.igem.org/mediawiki/2015/a/a3/Team-Brasil-USP_Pr-I13507.pdf">Pr_I13507</a>)<br><br> |
- Ligation <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, 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),<br><br> | - Ligation <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, 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),<br><br> | ||
-Transformation (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, Pl_I13507II, Pa_I13507II, Pr_I13507II, Pl_TermII, Pa_TermII, Pr_TermII)<br><br> | -Transformation (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, Pl_I13507II, Pa_I13507II, Pr_I13507II, Pl_TermII, Pa_TermII, Pr_TermII)<br><br> | ||
Line 1,232: | Line 1,245: | ||
-NotI digestion (Pl_I13507II, Pe_I13507II, Pl_TermII, Pa_TermII, Pr_TermII)<br><br> | -NotI digestion (Pl_I13507II, Pe_I13507II, Pl_TermII, Pa_TermII, Pr_TermII)<br><br> | ||
- SacI digestion again (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt)<br><br> | - SacI digestion again (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt)<br><br> | ||
− | -Agarose gel (Pl_I13507II, Pe_I13507II, Pl_TermII, Pa_TermII, Pr_TermII, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a> | + | -Agarose gel (Pl_I13507II, Pe_I13507II, Pl_TermII, Pa_TermII, Pr_TermII, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt)<br><br> |
− | -Ligation (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, | + | -Ligation (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, <a href="https://static.igem.org/mediawiki/2015/9/9b/Team-Brasil-USP_linkerGFP-termA.pdf">linkerGFP_termA</a> (J23101 + linkerGFP_Term), <a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a> (J23117 + linkerGFP_Term), <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a> (J23106 + linkerGFP_Term),<br><br> |
− | -Transformation (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, | + | -Transformation (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, <a href="https://static.igem.org/mediawiki/2015/9/9b/Team-Brasil-USP_linkerGFP-termA.pdf">linkerGFP_termA</a>, <a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a>, <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a><br><br> |
<br> | <br> | ||
Line 1,240: | Line 1,253: | ||
<br> | <br> | ||
- Transformation (TAT_ lcp_RBS,<a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_ lcp</a>,<a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_ roxA</a>, OmpA_roxA_Pt)<br><br> | - Transformation (TAT_ lcp_RBS,<a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_ lcp</a>,<a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_ roxA</a>, OmpA_roxA_Pt)<br><br> | ||
− | -Inoculum ( | + | -Inoculum (<a href="https://static.igem.org/mediawiki/2015/9/9b/Team-Brasil-USP_linkerGFP-termA.pdf">linkerGFP_termA</a>, <a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a>, <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a>)<br><br> |
<br> | <br> | ||
</td> | </td> | ||
Line 1,246: | Line 1,259: | ||
<td valign=top align=left> 12/09 | <td valign=top align=left> 12/09 | ||
<br> | <br> | ||
− | - Miniprep ( | + | - Miniprep (<a href="https://static.igem.org/mediawiki/2015/9/9b/Team-Brasil-USP_linkerGFP-termA.pdf">linkerGFP_termA</a>, <a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a>, <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a>)<br><br> |
− | - Not digestion ( | + | - Not digestion (<a href="https://static.igem.org/mediawiki/2015/9/9b/Team-Brasil-USP_linkerGFP-termA.pdf">linkerGFP_termA</a>, <a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a>, <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a>)<br><br> |
- Xbal and NotI digestion (RoxA, signalTAT and Lcp)<br><br> | - Xbal and NotI digestion (RoxA, signalTAT and Lcp)<br><br> | ||
− | -Agarose gel ( | + | -Agarose gel (<a href="https://static.igem.org/mediawiki/2015/9/9b/Team-Brasil-USP_linkerGFP-termA.pdf">linkerGFP_termA</a>, <a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a>, <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a>, RoxA, signalTAT and Lcp)<br><br> |
-Ligation <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a> (signalTAT + Lcp), <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_RoxA</a> (signalTAT + roxA)<br><br> | -Ligation <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a> (signalTAT + Lcp), <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_RoxA</a> (signalTAT + roxA)<br><br> | ||
<br> | <br> | ||
Line 1,268: | Line 1,281: | ||
- Agarose gel ( <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-Term.pdf">Pa_Term</a>, signalTAT, OmpA, Lcp, RoxA)<br><br> | - Agarose gel ( <a href="https://static.igem.org/mediawiki/2015/4/40/Team-Brasil-USP_Pa-Term.pdf">Pa_Term</a>, signalTAT, OmpA, Lcp, RoxA)<br><br> | ||
-Ligation (pSB1A2 + RoxA, pSB1A2 + Lcp, pSB1A2 + OmpA and pSB1A2 + SignalTAT)<br><br> | -Ligation (pSB1A2 + RoxA, pSB1A2 + Lcp, pSB1A2 + OmpA and pSB1A2 + SignalTAT)<br><br> | ||
− | -3A assembly (<a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_RoxA</a>(pSB1A2 + | + | -3A assembly (<a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_RoxA</a> (pSB1A2 + signalTAT + RoxA), <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a> (pSB1A2 + signalTAT + Lcp),<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_RoxA</a> (pSB1A2 +OmpA+RoxA), <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf"> OmpA_Lcp </a> (pSB1A2 + OmpA + Lcp)<br><br> |
− | -Transformation (RoxA, Lcp, OmpA, SignalTAT, (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> | + | -Transformation (RoxA, Lcp, OmpA, SignalTAT, (<a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> |
− | - Inoculum ( | + | - Inoculum (<a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a>, <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a>)<br><br> |
<br> | <br> | ||
Line 1,277: | Line 1,290: | ||
<br> | <br> | ||
- Ligation again (pSB1A2 + RoxA, pSB1A2 + Lcp, pSB1A2 + OmpA and pSB1A2 + SignalTAT)<br><br> | - Ligation again (pSB1A2 + RoxA, pSB1A2 + Lcp, pSB1A2 + OmpA and pSB1A2 + SignalTAT)<br><br> | ||
− | - 3A assembly again (<a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_RoxA</a>(pSB1A2 +signalTA+RoxA),<a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>(pSB1A2 +signalTA+Lcp), OmpA_RoxA(pSB1A2 +OmpA+RoxA),OmpA_Lcp( pSB1A2 +OmpA+Lcp))<br><br> | + | - 3A assembly again (<a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_RoxA</a>(pSB1A2 +signalTA + RoxA), <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>(pSB1A2 +signalTA + Lcp), OmpA_RoxA(pSB1A2 + OmpA + RoxA), OmpA_Lcp( pSB1A2 + OmpA + Lcp))<br><br> |
− | - Sample preparation ( | + | - Sample preparation (<a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a>, <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a>)<br><br> |
− | - Measurements ( | + | - Measurements (<a href="https://static.igem.org/mediawiki/2015/d/d9/Team-Brasil-USP_linkerGFP-termB.pdf">linkerGFP_termB</a>, <a href="https://static.igem.org/mediawiki/2015/d/d2/Team-Brasil-USP_linkerGFP-termC.pdf">linkerGFP_termC</a>)<br><br> |
<br> | <br> | ||
Line 1,295: | Line 1,308: | ||
<td valign=top align=left> 17/09 | <td valign=top align=left> 17/09 | ||
<br> | <br> | ||
− | - Inoculum (RoxA, Lcp, OmpA, SignalTAT, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a> | + | - Inoculum (RoxA, Lcp, OmpA, SignalTAT, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> |
- Cell lyses<br><br> | - Cell lyses<br><br> | ||
- Western blot (Lcp)<br><br> | - Western blot (Lcp)<br><br> | ||
Line 1,307: | Line 1,320: | ||
- Miniprep (RoxA, Lcp, OmpA, SignalTAT, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> | - Miniprep (RoxA, Lcp, OmpA, SignalTAT, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> | ||
- NotI digestion (RoxA, Lcp, OmpA, SignalTAT, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> | - NotI digestion (RoxA, Lcp, OmpA, SignalTAT, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> | ||
− | - Agarose gel (RoxA, Lcp, OmpA, SignalTAT, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a> | + | - Agarose gel (RoxA, Lcp, OmpA, SignalTAT, <a href="https://static.igem.org/mediawiki/2015/1/15/Team-Brasil-USP_OmpA-Lcp.pdf">OmpA_Lcp</a>,<a href="https://static.igem.org/mediawiki/2015/0/03/Team-Brasil-USP_OmpA-RoxA.pdf">OmpA_roxA</a>, <a href="https://static.igem.org/mediawiki/2015/7/74/Team-Brasil-USP_TAT-Lcp.pdf">TAT_Lcp</a>, <a href="https://static.igem.org/mediawiki/2015/6/6f/Team-Brasil-USP_TAT-RoxA.pdf">TAT_roxA</a>)<br><br> |
- Western blot (Lcp)<br><br> | - Western blot (Lcp)<br><br> | ||
<br> | <br> | ||
Line 1,314: | Line 1,327: | ||
+ | </td> | ||
+ | <td> | ||
</td> | </td> | ||
</table> | </table> | ||
− | |||
− | |||
<hr class="fancy-line" /> | <hr class="fancy-line" /> |
Latest revision as of 19:10, 18 September 2015
Notebook
Calendar
![](https://static.igem.org/mediawiki/2015/7/7d/Team-Brasil-USP_MainLogo.png)
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
| 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
- Transformation 6xHis_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_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_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_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_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, linkerGFP_termA (J23101 + linkerGFP_Term), linkerGFP_termB (J23117 + linkerGFP_Term), linkerGFP_termC (J23106 + linkerGFP_Term), -Transformation (OmpA_Lcp,OmpA_roxA, TAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, linkerGFP_termA, linkerGFP_termB, linkerGFP_termC | 11/09
- Transformation (TAT_ lcp_RBS,TAT_ lcp,TAT_ roxA, OmpA_roxA_Pt) -Inoculum (linkerGFP_termA, linkerGFP_termB, linkerGFP_termC) |
12/09
- Miniprep (linkerGFP_termA, linkerGFP_termB, linkerGFP_termC) - Not digestion (linkerGFP_termA, linkerGFP_termB, linkerGFP_termC) - Xbal and NotI digestion (RoxA, signalTAT and Lcp) -Agarose gel (linkerGFP_termA, linkerGFP_termB, linkerGFP_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 + signalTAT + RoxA), TAT_Lcp (pSB1A2 + signalTAT + 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_termB, linkerGFP_termC) | 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_termB, linkerGFP_termC) - Measurements (linkerGFP_termB, linkerGFP_termC) |
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_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_Lcp, TAT_roxA) - Western blot (Lcp) |