Team:Groningen/Notebook/Wetwork 12September2012

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Test GFP: reaction to rotten meat
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meat: 5 gram aliquot from freezer, 3 days at 20 degrees Celsius
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tested:
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Bacillus subtilis sp. 168 _ SboA_GFP in psac-cm (subjected to meat, and without meat) and WT (without meat)
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11:00 inoculate O/N culture in SMM in normal 37 degrees stove.
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*OD SboA = 0.302
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</head>
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*OD WT = 0.317
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12:30 SboA-GFP to micro-array test setup, start OD = 0.1 (see notebook Wetwork 02/09 and 27/06). Also put SboA-GFP and WT in normal 37 degrees stove (start OD = 0.1).
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<body><br><br><br>
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<p class="margin">
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Tom/Renske/Elbrich<br>
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<br>
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Test GFP: reaction to rotten meat<br>
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<br>
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meat: 5 gram aliquot from freezer, 3 days at 20 degrees Celsius<br>
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<br>
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tested:<br>
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Bacillus subtilis sp. 168 _ SboA_GFP in psac-cm (subjected to meat, and without meat) and WT (without meat)<br>
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<br>
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11:00 inoculate O/N culture in SMM in normal 37 degrees stove.
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<ul>
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<li>OD SboA = 0.302</li>
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<li>OD WT = 0.317</li>
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</ul>
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<p class="margin">12:30 SboA-GFP to micro-array test setup, start OD = 0.1 (see notebook Wetwork 02/09 and 27/06). Also put SboA-GFP and WT in normal 37 degrees stove (start OD = 0.1).<br>
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<br>
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<p class="margin">14:30 harvest cells
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<ul>
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<li>OD WT = 1.168</li>
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<li>OD SboA(no meat) = 1.023</li>
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<li>OD SboA(meat) = 1.153</li>
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</ul>
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<p class="margin">put cells on microscope slide for fluorescence microscopy:
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<ul><li>agarose (1% in SMM) 500 uL on slide</li>
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<li>spread with glass plate</li>
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<li>spot 1 uL of sample on slide (duplo)</li>
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</ul>
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<p class="margin">microscope results: no GFP, only autofluorescence.<br>
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<br><br>
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Renske<br>
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<br>
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put promoters of LMU Munchen in dh5a<br>
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<br>
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transformation of ~1 uL at 15:00<br>
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mistake! forgot to add LB medium!! --> Plate out anyway (amp plates) + O/N 5 mL culture of the rest..<br>
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<br>
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<br>
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O/N culture of: <br>
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<ul>
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<li>rfp-psac-cm in dh5a: 2x from 9/09 (our competent cells)</li>
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<li>rfp-psac-cm in dh5a: 5x from 10/09 (new competent cells)</li>
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</ul>
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<p class="margin">at 15:00, 37 degrees C.<br> <br>
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Emeraldo <br> <br>
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Another batch of competent cells in the making! <br> <br>
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Nisa <br> <br>
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Another cloning for pSac-Cm+promoters+reporters and BBa_K823023+promoters+reporters in progress. We borrowed some tubes of E. coli DH5a competent cell from Sjoerd, our lovely supervisor.  
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14:30 harvest cells
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<br>
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*OD WT = 1.168
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<A HREF="https://2012.igem.org/Team:Groningen/Notebook"><FONT COLOR=#ff6700>Back to notebook</FONT></A>
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*OD SboA(no meat) = 1.023
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</p><br><br>
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*OD SboA(meat) = 1.153
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</body>
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put cells on microscope slide for fluorescence microscopy:
+
-
* agarose (1% in SMM) 500 uL on slide
+
-
* spread with glass plate
+
-
* spot 1 uL of sample on slide (duplo)
+
-
 
+
-
microscope results: no GFP, only autofluorescence.
+
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<html>
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<A HREF="https://2012.igem.org/Team:Groningen/Notebook"><FONT COLOR=#ff6700>Back to notebook</FONT></A>
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</html>
</html>
{{Template:SponsorsGroningen2012}}
{{Template:SponsorsGroningen2012}}

Latest revision as of 14:59, 26 September 2012







Tom/Renske/Elbrich

Test GFP: reaction to rotten meat

meat: 5 gram aliquot from freezer, 3 days at 20 degrees Celsius

tested:
Bacillus subtilis sp. 168 _ SboA_GFP in psac-cm (subjected to meat, and without meat) and WT (without meat)

11:00 inoculate O/N culture in SMM in normal 37 degrees stove.

  • OD SboA = 0.302
  • OD WT = 0.317

12:30 SboA-GFP to micro-array test setup, start OD = 0.1 (see notebook Wetwork 02/09 and 27/06). Also put SboA-GFP and WT in normal 37 degrees stove (start OD = 0.1).

14:30 harvest cells

  • OD WT = 1.168
  • OD SboA(no meat) = 1.023
  • OD SboA(meat) = 1.153

put cells on microscope slide for fluorescence microscopy:

  • agarose (1% in SMM) 500 uL on slide
  • spread with glass plate
  • spot 1 uL of sample on slide (duplo)

microscope results: no GFP, only autofluorescence.


Renske

put promoters of LMU Munchen in dh5a

transformation of ~1 uL at 15:00
mistake! forgot to add LB medium!! --> Plate out anyway (amp plates) + O/N 5 mL culture of the rest..


O/N culture of:

  • rfp-psac-cm in dh5a: 2x from 9/09 (our competent cells)
  • rfp-psac-cm in dh5a: 5x from 10/09 (new competent cells)

at 15:00, 37 degrees C.

Emeraldo

Another batch of competent cells in the making!

Nisa

Another cloning for pSac-Cm+promoters+reporters and BBa_K823023+promoters+reporters in progress. We borrowed some tubes of E. coli DH5a competent cell from Sjoerd, our lovely supervisor.
Back to notebook



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