Team:SDU-Denmark/project-p

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(Difference between revisions)
(Motility assay)
(Flagella staining)
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When the E.coli strain MG1655 have a constitutive active transcription of the FlhDC operon more flagella’s were expected. The idée was to get a quantities measure of the increase flagella production by comparing the observed number flagellas on the wildtype MG1655 and the hyperflagellated MG1655. We tried to stain the flagella using silver staining and afterwards examined the bacteria under the microscope. We started the staining procedure before we had the mutated FlhDC operon as a composed part because we then had time to optimize the staining protocol and become really good at it. We started with staining DH5α and H10407.  A E-coli strain which do not express flagellas (negative controle) and a hyperflagellated E-coli (positive control), respectively. The bacteria were grown on agar plates ON and stained with this protocol. <br> <br>
When the E.coli strain MG1655 have a constitutive active transcription of the FlhDC operon more flagella’s were expected. The idée was to get a quantities measure of the increase flagella production by comparing the observed number flagellas on the wildtype MG1655 and the hyperflagellated MG1655. We tried to stain the flagella using silver staining and afterwards examined the bacteria under the microscope. We started the staining procedure before we had the mutated FlhDC operon as a composed part because we then had time to optimize the staining protocol and become really good at it. We started with staining DH5α and H10407.  A E-coli strain which do not express flagellas (negative controle) and a hyperflagellated E-coli (positive control), respectively. The bacteria were grown on agar plates ON and stained with this protocol. <br> <br>
Unfortunately it was not possible to see clear and significant different between the positive and negative control. Though at some placeses it could look like flagellas on the positive control but it was never enough to determine a different. After repeating the protocol four times each time with many samples we decided to give up on the protocol. <br>
Unfortunately it was not possible to see clear and significant different between the positive and negative control. Though at some placeses it could look like flagellas on the positive control but it was never enough to determine a different. After repeating the protocol four times each time with many samples we decided to give up on the protocol. <br>
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[[Image:Team_SDU-Denmark_Silver_staining_of_H10407_a_positive_control..JPG|550px|
=== Scanning Electron microscopy ===
=== Scanning Electron microscopy ===

Revision as of 22:25, 25 October 2010