Team:Warsaw/Calendar-Main/23 July 2008

From 2008.igem.org

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<p>Ligations <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">transformed</a> into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB + ampicillin.</p>
<p>Ligations <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">transformed</a> into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB + ampicillin.</p>
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<h3>Cloning of protein A DNA to OmpA constructs</h3><h4>Michał K.</h4>
 
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<ol>
 
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation of plasmids</a> from cultures inocluated on previous day (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-alpha>pACYC177+OmpA_A_alpha</a>). </li>
 
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<li>Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids with BamHI and SacI (BamHI buffer).</li><li> Gel electrophoresis - proper clone found. (Fig. 1.)</li></ol>
 
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</p>
 
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<p>
Setup of overnight culture <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> (LB broth) for preparation of chemocompetent bacteria.</p>
Setup of overnight culture <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> (LB broth) for preparation of chemocompetent bacteria.</p>
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<h3>Cloning of protein A DNA to OmpA constructs</h3><h4>Michał K.</h4>
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<ol>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation of plasmids</a> from cultures inocluated on previous day (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-alpha>pACYC177+OmpA_A_alpha</a>). </li>
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<li>Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids with BamHI and SacI (BamHI buffer).</li><li> Gel electrophoresis - proper clone found. (Fig. 1.)</li></ol>
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</p>
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<img src="https://static.igem.org/mediawiki/2008/f/fb/23_july.jpg" width=300/> <var><b>Fig. 1. Control SacI/BamHI digests of pACYC177+OmpA_A_alpha</b><br>  
<img src="https://static.igem.org/mediawiki/2008/f/fb/23_july.jpg" width=300/> <var><b>Fig. 1. Control SacI/BamHI digests of pACYC177+OmpA_A_alpha</b><br>  

Revision as of 17:17, 26 October 2008

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Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin

  1. Isolation of plasmids from transformants.
  2. Digest of plasmids with SacI and NotI (BamHI buffer).
  3. Gel electrophoresis of products of digest.
  4. 2 selected clones were send to DNA sequencing lab.

Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA

Paweł

Ligations transformed into TOP10 and plated on LB + ampicillin.

Antoni

Setup of overnight culture E. coli TOP10 (LB broth) for preparation of chemocompetent bacteria.

Cloning of protein A DNA to OmpA constructs

Michał K.

  1. Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_A_alpha).
  2. Control digest of isolated plasmids with BamHI and SacI (BamHI buffer).
  3. Gel electrophoresis - proper clone found. (Fig. 1.)

Fig. 1. Control SacI/BamHI digests of pACYC177+OmpA_A_alpha
1-4. digested plasmids isolated from transformants obtained previous day
5. Marker


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