Paris/July 12

From 2007.igem.org

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We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:
We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:
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* left : Acinetobacter grown in Low Nitrogen Mineral medium (see Protocol)
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Visualisation of colonies stained by Nile Red. It seems that only cells in the middle of colonies (i.e. "old" cells) are producing triglycerides whereas cells on the edges doesn't. <br>
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* right : Acinetobacter grow in Low Nitrogen Mineral medium with Nile Red (see Protocol).
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[[Image:Paris_Acineto_Colonies.jpg|500px|center]]]
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Exposure time and scaling is the same for both images. Clearly, some cells on the right are stained by Nile Red, which is specific to triglycerides, whereas almost all cells on the left are not stained. Tipically, the size of one cell is about 5µm.
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[[Image:Paris_Acineto_NileRed.jpg|850px]]
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Revision as of 14:46, 25 July 2007

Contents

Preparation of a stock of phage on w121:

David B:

  • Filtration of the Cacl2 and MgSO4 stocks
  • Launch a culture:
    • 15ml LB
    • 225µl Cacl2
    • 450µl MgSO4
    • 90µl DAP
    • 150µl W121 from an ON culture
  • Let grow until OD=0.2 (~1h)
  • Add 100µl P1 (stock 1010/ml)
  • Incubate 2h
  • Add 200µl Chloroform
  • Vortex, centrifuge (15min @ 3000rpm)
  • Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C

Titration of bacteriophages P1

Nicolas C.
We measure the strength of former P1 preparation :

  • The stock prepared the 3.7.7
  • The stock prepared the 8.7.7
  • The stock prepared today (12.7.7)

See results.
See protocols for details.

Culture of FtsZ TS

David B
We want to isolate a good clone of the strain FstZ TS
From the plate of FtsZ TS84 121:
isolation of 6 clones of the 121.1 on LBA+tet incubated at 42°C + Culture ON at 30°C
If one clone do not grow at 42, we'll use it to do the transduction.
See result.

Acinetobacter microscopy

We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days: Visualisation of colonies stained by Nile Red. It seems that only cells in the middle of colonies (i.e. "old" cells) are producing triglycerides whereas cells on the edges doesn't.

Paris Acineto Colonies.jpg
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