Team:Goettingen/week7-1

From 2012.igem.org

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<ul>
<ul>
<li>Experiment: <br>
<li>Experiment: <br>
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Two strains with different promotor strengths were mixed 1:1 (OD600) and dropped onto a trypton plate. After several hours of swimming, the bacteria from the most outer edge of the swimming halo were picked and quantified to identify the ratio between the both starter-strains.</li><br>
+
Two strains with different promotor strengths were mixed 1:1 (OD600) and dropped onto a trypton plate. After several hours of swimming, the bacteria from the most outer edge of the swimming halo were picked and quantified to identify the ratio between the both starter-strains.</li>
<li>Observations & Results: <br>
<li>Observations & Results: <br>
On each quater of a 150 mm petri dish was one bacteria colony droped. Therefore, the swimming ability of all four strains could be compared directly.
On each quater of a 150 mm petri dish was one bacteria colony droped. Therefore, the swimming ability of all four strains could be compared directly.
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<ul>
<ul>
<li>Experiment: <br>
<li>Experiment: <br>
-
On minimal plates (only 0.3% agar and 0.5% NaCl) was one strain dropped. In a distance of about 2 cm was an attractant dropped, to induce directed chemotaxis.</li><br>
+
On minimal plates (only 0.3% agar and 0.5% NaCl) was one strain dropped. In a distance of about 2 cm was an attractant dropped, to induce directed chemotaxis.</li>
<li>Observations & Results: <br>
<li>Observations & Results: <br>
No directed swimming or swimming at all was detectable. The aspartat attractant might not have reached the bacteria.
No directed swimming or swimming at all was detectable. The aspartat attractant might not have reached the bacteria.
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<h2><b>V06_14<b></h2><br>
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<b> Directed swimming with gradient plates</b><br>
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<ul>
 +
<li>Experiment: <br>
 +
To get a trypton gradient into the plates, they were poured in two steps. In the first step, the plate were shifted to a certain grad, to get more agar on one side to plate. In the second step, after hardening of the agar from step one, the plate is put back to normal and the other half is filled with agar. This all leads to plates, that have a linear gradient of chemoatractant in the agar. The strains from V06_11 are dropped on these plates to induce them to perform directed swimming.</li>
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</ul>
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<li>Observations & results:<br>
 +
No directed swimming was detectable
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</ul>
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<br></td></tr>
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</table>
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<table cellpadding="20 px" border="1" bordercolor="black" valign="top">
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<tr bordercolor="black" valign="top">
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<td width="900" bordercolor="black" valign="top">
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<h2><b>V06_15<b></h2><br>
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<b> Preparation of diverse agar compositions</b><br>
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<ul>
 +
<li>Experiment: <br>
 +
Four different agar compositions were produced: 1) 2x 400 mL: 1% trypton, 0.5% NaCl, 0.3% agar; 2) 2x 400 mL: 0.5% trypton, 0.5% NaCl, 0.3% agar; 3) 3x 400 mL: 0.5% NaCl, 0.3% agar; 4) 1x 400 mL: 0.5% NaCl, 0.3% agar, 0.25% aspartate.</li>
 +
</ul>
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<li>Observations & results:<br>
 +
No directed swimming was detectable
 +
</ul>
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<br></td></tr>
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</table>
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Revision as of 12:49, 26 September 2012

Deutsch  / English 

#1 Selection / Swimming - 7th week

Back to overview

V06_11


Swimming assay with trypton agar
  • Experiment:
    A variety of agar conditions were tested: 1) 1% trypton, 0.3% agar, 0.5% NaCl; 2) 0.5% trypton, 0.3%, 0.5% NaCl; 3) like 1, but with additionally 0.5% glcose; 4) like 2), but with additionally 0.5% glucose. The swimming assay was performed as mentioned in methods.
  • Observations & results:
    All plates w/o glucose showed strong swimming over night. Therefore, glucose could be considered to be taxis-inhbiting.

V06_12


Comined swimming assays
  • Experiment:
    The strains from the previos experiments were combined on a single plate (several distinct drops on the same plate) to compare their swimming ability directly.

V06_13


V06_13_1: Separation assay
  • Experiment:
    Two strains with different promotor strengths were mixed 1:1 (OD600) and dropped onto a trypton plate. After several hours of swimming, the bacteria from the most outer edge of the swimming halo were picked and quantified to identify the ratio between the both starter-strains.
  • Observations & Results:
    The plated did not show enough swimming, to ensure a positiv separation of the two strains. Further incubation times might be helpful.

V06_13_2: Comparing swimming assays
  • Experiment:
    Two strains with different promotor strengths were mixed 1:1 (OD600) and dropped onto a trypton plate. After several hours of swimming, the bacteria from the most outer edge of the swimming halo were picked and quantified to identify the ratio between the both starter-strains.
  • Observations & Results:
    On each quater of a 150 mm petri dish was one bacteria colony droped. Therefore, the swimming ability of all four strains could be compared directly.

V06_13_3: Chemotaxis assay with attractant
  • Experiment:
    On minimal plates (only 0.3% agar and 0.5% NaCl) was one strain dropped. In a distance of about 2 cm was an attractant dropped, to induce directed chemotaxis.
  • Observations & Results:
    No directed swimming or swimming at all was detectable. The aspartat attractant might not have reached the bacteria.

V06_14


Directed swimming with gradient plates
  • Experiment:
    To get a trypton gradient into the plates, they were poured in two steps. In the first step, the plate were shifted to a certain grad, to get more agar on one side to plate. In the second step, after hardening of the agar from step one, the plate is put back to normal and the other half is filled with agar. This all leads to plates, that have a linear gradient of chemoatractant in the agar. The strains from V06_11 are dropped on these plates to induce them to perform directed swimming.
  • Observations & results:
    No directed swimming was detectable

    • V06_15


    Preparation of diverse agar compositions
    • Experiment:
      Four different agar compositions were produced: 1) 2x 400 mL: 1% trypton, 0.5% NaCl, 0.3% agar; 2) 2x 400 mL: 0.5% trypton, 0.5% NaCl, 0.3% agar; 3) 3x 400 mL: 0.5% NaCl, 0.3% agar; 4) 1x 400 mL: 0.5% NaCl, 0.3% agar, 0.25% aspartate.
  • Observations & results:
    No directed swimming was detectable

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