Paris/July 12
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(→Culture of FtsZ TS) |
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From the plate of FtsZ TS84 121: <br> | From the plate of FtsZ TS84 121: <br> | ||
isolation of 6 clones of the 121.1 on LBA+tet incubated at 42°C + Culture ON at 30°C | 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 | ||
Revision as of 14:26, 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
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
If one clone do not grow at 42, we'll use it to do the transduction
