Team:Warsaw/Calendar-Main/18 June 2008

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Preparation of constructs with OmpA protein fusions

Michał K.

Isolation of pACYC177.
Digest of purified pACYC177 with NdeI and BamHI (Tango 2x buffer).
Gel electrophoresis and isolation of proper band (3200 bp).
Clean-up of OmpA_alpha and OmpA_omega digest products.
Ligation of purified DNA (pACYC177 with OmpA_alpha and OmpA_omega DNA fragments).
Transformation of E.coli TOP10 with ligation products.
Plating transformants on LB+tetracycline.

Blue/white and rifampicin test

Michał L., Ewa, Marcin

Counting of blue colonies:

Strain	% of blue colonies
pMPMT5	99%
pMPMT5-AID	93%
pMPMT5-AIDT7	81%
pMPMT5-AID+T7	1%

Results of rifampicin test
Strain	Inducer / repressor	Number of colonies on rifampicin
Top10	none	8
Top10 pMPM-AID	0.1% Arabinose	72
Top10 pMPM-AID	none	4
Top10 pMPM-A+T13	0.1% Arabinose	21
Top10 pMPM-A+T13	none	6
Top10 pMPM-AT6	0.1% Arabinose	148
Top10 pMPM-AT6	none	27

@@ Line 11: / Line 11: @@
 <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E.coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligation products.</li>
 <li>Plating transformants on LB+tetracycline.</li></ol>
 <h3> Blue/white and rifampicin test </h3>
 <h4>Michał L., Ewa, Marcin</h4>
-<p>Counting of blue colonies. Same results as on <a href=https://2008.igem.org/Team:Warsaw/Calendar-Main/12_June_2008>12/06/2008</a>. </p>
+<p>Counting of blue colonies: </p>
+<table id="result" align="left">
+<tr><th>Strain</th><th>% of blue colonies</th></tr>
+<tr><td>pMPMT5</td><td>99%</td></tr>
+<tr><td>pMPMT5-AID</td><td>93%</td></tr>
+<tr><td>pMPMT5-AIDT7</td><td>81%</td></tr>
+<tr><td>pMPMT5-AID+T7</td><td>1%</td></tr>
+</table>
+<table class="month" align="left" width="100%"><tr><td><br/></td></tr></table>
+<table id="result" align="left">
+<tr><th colspan="3"> Results of rifampicin test</th></tr>
+<tr><th>Strain</th><th>Inducer / repressor</th><th>Number of colonies on rifampicin</th></tr>
+<tr><th>Top10</th><td>none</td><td>8</td></tr>
+<tr><th>Top10 pMPM-AID</th><td>0.1% Arabinose</td><td>72</td></tr>
+<tr><th>Top10 pMPM-AID</th><td>none</td><td>4</td></tr>
+<tr><th>Top10 pMPM-A+T13</th><td>0.1% Arabinose</td><td>21</td></tr>
+<tr><th>Top10 pMPM-A+T13</th><td>none</td><td>6</td></tr>
+<tr><th>Top10 pMPM-AT6</th><td>0.1% Arabinose</td><td>148</td></tr>
+<tr><th>Top10 pMPM-AT6</th><td>none</td><td>27</td></tr>
+</table>
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