Team:Marburg:Project:Notebook:May
From 2014.igem.org
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<fieldset class="exp15"> | <fieldset class="exp15"> | ||
- | <legend><a name="exp15.22">15.22 Restriction of 3 | + | <legend><a name="exp15.22">15.22 Restriction of 3 isolated plasmids (6.5.14) with <i>Nco</i>I and <i>Bam</i>HI</a></legend> |
<div class="aim"> | <div class="aim"> | ||
<p>Aim: Test whether the isolated plasmid contains the 2 kb insert (hag-flank construct).</p> | <p>Aim: Test whether the isolated plasmid contains the 2 kb insert (hag-flank construct).</p> | ||
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</fieldset> | </fieldset> | ||
<fieldset class="exp17"> | <fieldset class="exp17"> | ||
- | <legend><a name="exp17.3">17.3 First Plate Reader Measurement with <i>Bacillus subtilis</i> wildtype and strain 186 | + | <legend><a name="exp17.3">17.3 First Plate Reader Measurement with <i>Bacillus subtilis</i> wildtype and strain 186 containing a <i>gfp</i></a></legend> |
<div class="aim"> | <div class="aim"> | ||
<p>Aim: Test for differences between the two strains and first check if experiment works at all</p> | <p>Aim: Test for differences between the two strains and first check if experiment works at all</p> | ||
</div> | </div> | ||
+ | <br /> | ||
+ | <img src="https://static.igem.org/mediawiki/2014/3/3b/MR_20140520_MTA_growth_Bsubtilis_wt_gfp_MM_LB.jpg" width="40%" /> | ||
+ | <br /> | ||
+ | <img src="https://static.igem.org/mediawiki/2014/0/09/MR_20140520_MTA_fluorescence_bsubtilis_wt_gfp_MM_xyl_LB.jpg" width="40%" /> | ||
+ | <br /> | ||
+ | <p>Both strains grew best in LB medium like expected, a clearly better growth was shown by the wildtype in the minimal medium compared to the <i>gfp</i> bearing strain.</p> | ||
+ | <p>The fluorescence is shown in dependence of the culture growth in order to relativise the fluorescence signal. Like expected, the wt exhibited no fluorescence in both media. The fluorescence of the <i>gfp</i> strain was higher in the minimal medium with xylose, than without xylose.</p> | ||
</fieldset> | </fieldset> | ||
</div> | </div> | ||
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<legend><a name="exp17.6">17.6 MTA Bacillus Nose Ag </a></legend> | <legend><a name="exp17.6">17.6 MTA Bacillus Nose Ag </a></legend> | ||
<div class="exp-content"> | <div class="exp-content"> | ||
- | <p> | + | <p>Transformation from 20.5.14: one colony on the Ag Nose plate<br /> |
- | Inoculation of shaking culture in the morning together with the wildtype and the gfp strain | + | Inoculation of shaking culture in the morning together with the wildtype and the <i>gfp</i> strain in LB liquid without antibiotics</p> |
- | <p> | + | <p>Inoculate 96 well plate in the afternoon 15:00</p> |
<p>Prepare Cu and Ag solutions to test different concentrations in MTA and see if the Ag nose responds to Ag and also if it is specific for Ag and does not respond to Cu</p> | <p>Prepare Cu and Ag solutions to test different concentrations in MTA and see if the Ag nose responds to Ag and also if it is specific for Ag and does not respond to Cu</p> | ||
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<p>Ag and Cu in stock solution of 10 mM have been prepared<br /> | <p>Ag and Cu in stock solution of 10 mM have been prepared<br /> | ||
Added (Concentration): 11 mM, 100µM and 10µM<br /> | Added (Concentration): 11 mM, 100µM and 10µM<br /> | ||
- | Cells: Ag Nose mutant, wildtype (AG GM) and gfp | + | Cells: Ag Nose mutant, wildtype (AG GM) and <i>gfp</i></p> |
+ | <br /> | ||
+ | <p>The MTA showed no significant increase of fluorescence. The bacteria had difficulties in growth, thus it seemed that the metal ion concentrations were too high.</p> | ||
</div> | </div> | ||
</fieldset> | </fieldset> | ||
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</tr> | </tr> | ||
<tr> | <tr> | ||
- | <th scope="row"><strong>Template | + | <th scope="row"><strong>Template ~10µg</strong></th> |
<td>15</td> | <td>15</td> | ||
<td>20</td> | <td>20</td> |
Latest revision as of 19:28, 16 October 2014
Notebook: May