Paris/July 12

From 2007.igem.org

(Difference between revisions)
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David B:
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==Preparation of a stock of phage on w121:==
==Preparation of a stock of phage on w121:==
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David B: 
* Filtration of the Cacl2 and MgSO4 stocks
* Filtration of the Cacl2 and MgSO4 stocks
* Launch a culture:
* Launch a culture:
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* Let grow until OD=0.2 (~1h)
* Let grow until OD=0.2 (~1h)
* Add 100µl P1 (stock 10^10/ml)
* Add 100µl P1 (stock 10^10/ml)
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* Incubate 2h
 
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* Add 1ml of the ON culture of W121
 
* Incubate 2h
* Incubate 2h
* Add 200µl Chloroform
* Add 200µl Chloroform
* Vortex, centrifuge (15min @ 3000rpm)
* Vortex, centrifuge (15min @ 3000rpm)
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* Recover the supernatant = Stock keep at 4°C
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* Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C
[[User:Nicolas C.|Nicolas C.]]
[[User:Nicolas C.|Nicolas C.]]
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* The stock prepared the 8.7.7
* The stock prepared the 8.7.7
See protocols for details.
See protocols for details.
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== Culture of FtsZ TS ==
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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

Revision as of 14:00, 12 July 2007


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 10^10/ml)
  • Incubate 2h
  • Add 200µl Chloroform
  • Vortex, centrifuge (15min @ 3000rpm)
  • Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C

Nicolas C.

Titration of bacteriophages P1

We measure the strength of former P1 preparation :

  • The stock prepared the 3.7.7
  • The stock prepared the 8.7.7

See protocols for details.

Culture of FtsZ TS

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