Team:Warsaw/Calendar-Main/27 September 2008

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<h3>Mutagenesis of protein A</h3><h4>Paweł</h4>
<h3>Mutagenesis of protein A</h3><h4>Paweł</h4>
-
<p><ol><li>Isolation of plasmid DNA from overnight cultures (it was really tough work...).</li>
+
<p><ol><li>Isolation of plasmid DNA from overnight cultures (it was really hard work...).</li>
<li>Control digest of isolated plasmids with KpnI and BamHI (if mutagenesis really occured, ~550bp band should be visible).</li>
<li>Control digest of isolated plasmids with KpnI and BamHI (if mutagenesis really occured, ~550bp band should be visible).</li>
<li>Results of digest:</li>
<li>Results of digest:</li>
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<li>ADelL+KpnI and ADelP primers mutagenesis:</li>
<li>ADelL+KpnI and ADelP primers mutagenesis:</li>
<ul>
<ul>
-
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYClac+ompA-A-omega</a>: <b>0</b> positive, <b>5</b> negative</li>
+
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BOmpA-deltaA-omega>pACYClac+ompA-&Delta;A-omega</a>: <b>0</b> positive, <b>5</b> negative</li>
-
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-alpha>pACYClac+ompa-A-alpha</a>: <b>3</b> positive, <b>7</b> negative</li>
+
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>pACYClac+ompa-&Delta;A-alpha</a>: <b>3</b> positive, <b>7</b> negative</li>
-
<li>pACYClac+ ompa-alpha-A: <b>1</b> positive, <b>2</b> negative</li>
+
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-deltaA>pACYClac+OmpA-omega-&Delta;A</a>: <b>1</b> positive, <b>2</b> negative</li>
</ul>
</ul>
<li>AMutL+KpnI and AMutP mutagenesis:</li>
<li>AMutL+KpnI and AMutP mutagenesis:</li>
<ul>
<ul>
-
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYClac+ompA-A-omega</a>: <b>4</b> positive, <b>16</b> negative</li>
+
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BOmpA-deltaA-omega>pACYClac+ompA-&Delta;A-omega</a>: <b>4</b> positive, <b>16</b> negative</li>
-
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-alpha>pACYClac+ompa-A-alpha</a>: <b>4</b> positive, <b>8</b> negative</li>
+
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>pACYClac+ompa-&Delta;A-alpha</a>: <b>4</b> positive, <b>8</b> negative</li>
-
<li>pACYClac+ ompa-alpha-A: <b>2</b> positive, <b>15</b> negative</li>
+
<li><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-deltaA>pACYClac+OmpA-omega-&Delta;A</a>: <b>2</b> positive, <b>15</b> negative</li>
</ul>
</ul>
</ul>  
</ul>  

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Mutagenesis of protein A

Paweł

  1. Isolation of plasmid DNA from overnight cultures (it was really hard work...).
  2. Control digest of isolated plasmids with KpnI and BamHI (if mutagenesis really occured, ~550bp band should be visible).
  3. Results of digest:
  4. Plasmids which gave positive digest results prepared for sequencing (one sample from each variant which gave positive result).

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