Team:Goettingen/week12-2
From 2012.igem.org
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<li>Experiment: <br> | <li>Experiment: <br> | ||
In order to test the influence of the different genes on the motility of the distinct strains over night cultures were prepared. On that account it is important to shake the cultures only at 140 rpm to spare the flagella. <br></li> | In order to test the influence of the different genes on the motility of the distinct strains over night cultures were prepared. On that account it is important to shake the cultures only at 140 rpm to spare the flagella. <br></li> | ||
+ | <li>Observations & Results: <br> | ||
+ | Not all cultures grew properly. Especially, those of <i>E.coli</i> BL21 featured a low cell density.</li> | ||
<br></td></tr> | <br></td></tr> | ||
</table> | </table> | ||
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<table cellpadding="20 px" border="1" bordercolor="black" valign="top"> | <table cellpadding="20 px" border="1" bordercolor="black" valign="top"> | ||
<tr bordercolor="black" valign="top"> | <tr bordercolor="black" valign="top"> | ||
<td width="900" bordercolor="black" valign="top"> | <td width="900" bordercolor="black" valign="top"> | ||
- | <h2><b> | + | <h2><b>V07_18 </b></h2><br> |
- | <b> | + | <b> Performance of motility assay</b><br> |
<ul> | <ul> | ||
<li>Experiment: <br> | <li>Experiment: <br> | ||
- | + | LB medium containing ampicillin was inoculated with the over night culture and shaken until an optical density of 0.6 was reached. Meanwhile ampicillin containing M9 agar plates were prepared and autoclaved Whatman Filter Papers were imbued with tryptone solution. Then the cultures were dropped on the agar at a distance of 1 cm to the paper. On each plate also colonies hosting the empty vector pUC18 were dropped in order to have a reference. Each plate was prepared in duplicates.</li> | |
<li>Observations & Results: <br> | <li>Observations & Results: <br> | ||
- | The | + | The following freshly inoculated cultures did not grow very well and only reached an OD between 0.3 and 0.4 : |
+ | <i>fliC</i> (DH10B) in DH10B <br> | ||
+ | <i>fliC</i> (DH10B) in DH5alpha <br> | ||
+ | <i>fliC</i> (DH10B) in XL1 Blue <br> | ||
+ | <i>motB</i> in DH5alpha <br> | ||
+ | <i>motB</i> in XL1 Blue <br> | ||
+ | At the following day (19th of July) the colonies still looked exactly the same. Neither chemotaxis nor swimming could observed. | ||
+ | Two days after preparation of the swimming assay (20th of July) plates slight chemotaxis and motility could be perceived. <i>motB</i> and <i>fliC</i> overexpression seems to have a positive influence on bacterial motility in all strains. | ||
+ | At the 23rd of July quite big halos had formed. <i>motB</i> shows a strong effect, especially in DH10B. <i>fliC</i> (DH10B) seems to increase motility in all strains. However, the difference to the reference is not very drastic at all, but since the genes are not control under strong promoters this results seems reasonable. | ||
<br></td></tr> | <br></td></tr> | ||
</table> | </table> | ||
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<br> | <br> |
Revision as of 17:46, 16 September 2012
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