Team:Goettingen/week14-2

V07_30

Preparation of over night cultures

• Experiment:
  In order to perform a further swimming assay new over night cultures of fliC (DH10B), fliC (Salmonella), motA, motB and yhjH in DH10B, BL21, DH5α and XL1 Blue were prepared.
  


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V08_01 Preparation of over night cultures Experiment: Since some problems with the growing of the cultures appeared new over night cultures of fliC (DH10B), fliC (Salmonella), motA, motB and yhjH in DH10B, BL21, DH5α and XL1 Blue were prepared. </td></tr> </table> V08_02 V08_02_1 Performance of motility assay Experiment: LB medium containing ampicillin was inoculated with the over night culture and shaken until an optical density of 0.4-0.6 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. Two drops were rather applied into the agar than onto as done before, because some paper predicted faster motility. Also this time each plate was prepared in duplicates. Observations & Results: Unfortunately, even after a week no swimming could be observed only growth. We assume that the medium might not provide adequate swimming conditions and therefore we plan to test more motile strains and alter the agar composition if necessary. V08_02_2 Preparation of over night cultures Experiment: Since we recently received new E. coli strains that are more motile than our usually used laboratory stains new over night cultures of MG1655 and RP437 were prepared. V08_03 Performance of motility assay Experiment: This time the over night cultures were directly dropped to the swimming plates. In oder to test whether the medium or indeed our strains are responsible for the immobility of the bacteria the habitual M9 agar plates were prepared. However not antibiotics were used since the new strains are not equipped with any resistance yet. Then the autoclaved Whatman Filter Papers were imbued with tryptone solution were placed in the middle of the plate and the cultures were dropped on the agar at a distance of 1 cm to the paper. Also this time each plate was prepared in duplicates. Observations & Results: The following day (4th of August) no motility could be observed. Since both strains are known to move very quick and are supposed to swarm the whole plate during hours we suggest that the medium composition is indeed not suitable. However, two days after preparation of the swimming assay (5th of August) some evidence for incipient swimming was visible, which grew to small halos the following days. Nevertheless, the slow progress confirmed our resolution to reconsider the composition of the plates. </i> </td></tr> </table> <a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a> </td> </font></td></tr></tbody></table></div> </td></tr> </table> </body> </html> {{GoettingenFooter}