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Team:Warsaw/Calendar-Main/2 October 2008
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primers on colonies from plates with transformations <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> (annealing temperature - 55°C,45 s of elongation step). </li> | primers on colonies from plates with transformations <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> (annealing temperature - 55°C,45 s of elongation step). </li> | ||
| - | <li> Gel electrophoresis.</li> | + | <li> Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/2_October_2008#fig1">Fig. 1</a>).</li> |
<li> Confirmed transformant colonies inoculated to liquid LB with ampicillin.</li> | <li> Confirmed transformant colonies inoculated to liquid LB with ampicillin.</li> | ||
</ol> | </ol> | ||
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/9/92/Kolonijny_rfp_omp_03_10_2008.jpg"></a> | <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/9/92/Kolonijny_rfp_omp_03_10_2008.jpg"></a> | ||
| - | <var><b>Fig. | + | <var><b>Fig. 1.</b>Colony PCR with OmpaL_N and OmpaP_link. <br> |
| + | 1. Marker<br> | ||
| + | 4, 14, 19, 20, 24. Proper bands visible. <br> </var> | ||
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<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 plasmids from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103004>Z(BBa_K103004)</a>).</li> | <ol><li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103004>Z(BBa_K103004)</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 (<li> Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/2_October_2008#fig2">Fig. 2</a>).</li></ol> |
| - | <a name=" | + | <a name="fig2"><img src="https://static.igem.org/mediawiki/2008/6/6f/Traw_02_10_2008.jpg"></a> |
| - | <var><b>Fig. | + | <var><b>Fig. 2.</b>Control digest of pSB1A3+Z(BBa_K103004)<br> |
| + | 1. Marker<br> | ||
| + | 3-5. Clones carrying BBa_K103004</var> | ||
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<ol> | <ol> | ||
<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of <a href=http://partsregistry.org/Part:BBa_K103018>BBa_K103018</a> fragment with EcoRI and BcuI (BamHI buffer). </li> | <li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of <a href=http://partsregistry.org/Part:BBa_K103018>BBa_K103018</a> fragment with EcoRI and BcuI (BamHI buffer). </li> | ||
| - | <li>Gel electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 1200 bp. </li> | + | <li> Gel electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 1200 bp. <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/2_October_2008#fig3">Fig. 3</a>.</li> |
</ol> | </ol> | ||
| - | <a name=" | + | <a name="fig3"><img src="https://static.igem.org/mediawiki/2008/5/54/Go_02_10_2008_na_2_10.jpg"></a> |
| - | <var><b>Fig. | + | <var><b>Fig. 3.</b>Digest of OmpA_linker_omega_linker under Plac (BBa_K103018).<br> |
| + | 1. Marker<br> | ||
| + | 2. OmpA_linker_omega_linker under Plac (BBa_K103018)<br></var> | ||
Latest revision as of 20:34, 28 October 2008
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Preparation of linker_alpha (BBa_K103009)Michał K.Overnight ligation of isolated DNA fragments: pSB2K3 + linker_alpha (BBa_K103009). Preparation of linker_omega (BBa_K103013)Michał K.Overnight ligation of isolated DNA fragments: pSB2K3 + linker_omega (BBa_K103013). Preparation of OmpA-linker-omega-linker (BBa_K103016)Michał K.Overnight ligation of DNA fragments isolated on 30 September : pACYC177 + OmpA-linker-omega-linker (BBa_K103016). Preparation of OmpA-linker (BBa_K103006)Michał K.
Fig. 1.Colony PCR with OmpaL_N and OmpaP_link. 1. Marker 4, 14, 19, 20, 24. Proper bands visible. Preparation of Z(BBa_K103004)Michał K.
Fig. 2.Control digest of pSB1A3+Z(BBa_K103004)1. Marker 3-5. Clones carrying BBa_K103004 Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)Michał K.
Fig. 3.Digest of OmpA_linker_omega_linker under Plac (BBa_K103018).1. Marker 2. OmpA_linker_omega_linker under Plac (BBa_K103018)
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