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Team:Groningen/Notebook/Wetwork 12September2012
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Bacillus subtilis sp. 168 _ SboA_GFP in psac-cm (subjected to meat, and without meat) and WT (without meat)<br> | Bacillus subtilis sp. 168 _ SboA_GFP in psac-cm (subjected to meat, and without meat) and WT (without meat)<br> | ||
<br> | <br> | ||
| - | 11:00 inoculate O/N culture in SMM in normal 37 degrees stove. | + | 11:00 inoculate O/N culture in SMM in normal 37 degrees stove. |
<ul> | <ul> | ||
<li>OD SboA = 0.302</li> | <li>OD SboA = 0.302</li> | ||
<li>OD WT = 0.317</li> | <li>OD WT = 0.317</li> | ||
</ul> | </ul> | ||
| - | |||
<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> | <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> | ||
<br> | <br> | ||
| - | <p class="margin">14:30 harvest cells | + | <p class="margin">14:30 harvest cells |
<ul> | <ul> | ||
<li>OD WT = 1.168</li> | <li>OD WT = 1.168</li> | ||
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<li>OD SboA(meat) = 1.153</li> | <li>OD SboA(meat) = 1.153</li> | ||
</ul> | </ul> | ||
| - | + | <p class="margin">put cells on microscope slide for fluorescence microscopy: | |
| - | <p class="margin">put cells on microscope slide for fluorescence microscopy:< | + | <ul><li>agarose (1% in SMM) 500 uL on slide</li> |
| - | < | + | |
<li>spread with glass plate</li> | <li>spread with glass plate</li> | ||
<li>spot 1 uL of sample on slide (duplo)</li> | <li>spot 1 uL of sample on slide (duplo)</li> | ||
</ul> | </ul> | ||
| - | |||
<p class="margin">microscope results: no GFP, only autofluorescence.<br> | <p class="margin">microscope results: no GFP, only autofluorescence.<br> | ||
<br><br> | <br><br> | ||
Renske<br> | Renske<br> | ||
<br> | <br> | ||
| - | + | put promoters of LMU Munchen in dh5a<br> | |
<br> | <br> | ||
| - | + | transformation of ~1 uL at 15:00<br> | |
| - | + | mistake! forgot to add LB medium!! --> Plate out anyway (amp plates) + O/N 5 mL culture of the rest..<br> | |
<br> | <br> | ||
<br> | <br> | ||
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<li>rfp-psac-cm in dh5a: 5x from 10/09 (new competent cells)</li> | <li>rfp-psac-cm in dh5a: 5x from 10/09 (new competent cells)</li> | ||
</ul> | </ul> | ||
| - | <p class="margin">at 15:00, 37 degrees C.<br> | + | <p class="margin">at 15:00, 37 degrees C.<br> <br> |
| + | Emeraldo <br> <br> | ||
| + | Another batch of competent cells in the making! <br> <br> | ||
| + | Nisa <br> <br> | ||
| + | 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. | ||
<br> | <br> | ||
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.
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