Team:Warsaw/Calendar-Main/23 July 2008

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<h3>Preparation of chemocompetent bacteria</h3>
<h4>Antoni</h4>
<h4>Antoni</h4>
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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>  
<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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<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 (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/23_July_2008#fig1">Fig. 1.</a>).</li></ol>
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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>  
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/f/fb/23_july.jpg" width=300/></a> <var><b>Fig. 1. </b>Control SacI/BamHI digests of pACYC177+OmpA_A_alpha<br>  
1-4. digested plasmids isolated from transformants obtained previous day<br>
1-4. digested plasmids isolated from transformants obtained previous day<br>

Latest revision as of 21:23, 29 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.

Preparation of chemocompetent bacteria

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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