Team:Warsaw/Calendar-Main/3 October 2008
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- | <h3>Preparation of | + | <h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103009>linker_alpha (BBa_K103009)</a></h3> |
<h4>Piotr</h4> | <h4>Piotr</h4> | ||
<ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligation <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009)</a>.</li> | <ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligation <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009)</a>.</li> | ||
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- | <h3>Preparation of | + | <h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103013>linker_omega (BBa_K103013)</a></h3> |
<h4>Piotr</h4> | <h4>Piotr</h4> | ||
<ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligation <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103013>linker_omega (BBa_K103013)</a>.</li> | <ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligation <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103013>linker_omega (BBa_K103013)</a>.</li> | ||
<li> Plating on LB with kanamycin.</li></ol> | <li> Plating on LB with kanamycin.</li></ol> | ||
- | <h3>Preparation of | + | <h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103016>OmpA-linker-omega-linker (BBa_K103016)</a></h3> |
<h4>Piotr</h4> | <h4>Piotr</h4> | ||
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- | <h3>Preparation of <a href=http://partsregistry.org/ | + | <h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103006>OmpA-linker (BBa_K103006)</a></h3> |
<h4>Michał K.</h4> | <h4>Michał K.</h4> | ||
- | <ol><li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+ | + | <ol><li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/Part:BBa_K103006>OmpA-linker (BBa_K103006)</a>).</li> |
- | <li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.</li></ol> | + | <li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/3_October_2008#fig1">Fig. 1</a>).</li></ol> |
+ | <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/0/0b/Traw_petxba_omp_07_10_2008_na_3_10.jpg"></a> | ||
+ | <var><b>Fig. 1.</b>Control digest of pSB1A3+OmpA-linker(BBa_K103006)<br> | ||
+ | 1. Marker<br> | ||
+ | 2-5. EcoRI/PstI control digest pSB1A3 carrying OmpA-linker (BBa_K103006)</var> | ||
- | <h3>Preparation of | + | |
+ | |||
+ | <h3>Preparation of vector for pT7 constructs</h3> | ||
<h4>Michał K.</h4> | <h4>Michał K.</h4> | ||
<ol> | <ol> | ||
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> with XbaI (Tango buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment (3 hr). </li> | <li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> with XbaI (Tango buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment (3 hr). </li> | ||
- | <li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> | + | <li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> (with removed XbaI site) - 6500 bp). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/3_October_2008#fig2">Fig. 2</a>. </li> |
</ol> | </ol> | ||
+ | <a name="fig2"><img src="https://static.igem.org/mediawiki/2008/c/c9/Go_02_10_2008_na_03_10.jpg"></a> | ||
+ | <var><b>Fig. 2.</b>XbaI digest of pET15b+OmpA_omega <br> | ||
+ | 1. Marker<br> | ||
+ | 2. XbaI digest pET15b+OmpA_omega</var> | ||
Latest revision as of 20:43, 28 October 2008
Preparation of linker_alpha (BBa_K103009)Piotr
Preparation of linker_omega (BBa_K103013)Piotr
Preparation of OmpA-linker-omega-linker (BBa_K103016)Piotr
Preparation of OmpA-linker (BBa_K103006)Michał K.
1. Marker 2-5. EcoRI/PstI control digest pSB1A3 carrying OmpA-linker (BBa_K103006) Preparation of vector for pT7 constructsMichał K.
1. Marker 2. XbaI digest pET15b+OmpA_omega
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