Paris/July 12
From 2007.igem.org
(Difference between revisions)
Nicolas C. (Talk | contribs) (→Titration of bacteriophages P1) |
Nicolas C. (Talk | contribs) (→Preparation of a stock of phage on w121:) |
||
| Line 9: | Line 9: | ||
** 150µl W121 from an ON culture | ** 150µl W121 from an ON culture | ||
* Let grow until OD=0.2 (~1h) | * Let grow until OD=0.2 (~1h) | ||
| - | * Add 100µl P1 (stock 10 | + | * Add 100µl P1 (stock 10<sup>10</sup>/ml) |
* Incubate 2h | * Incubate 2h | ||
* Add 200µl Chloroform | * Add 200µl Chloroform | ||
* Vortex, centrifuge (15min @ 3000rpm) | * Vortex, centrifuge (15min @ 3000rpm) | ||
* Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C | * Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C | ||
| - | |||
==Titration of bacteriophages P1 == | ==Titration of bacteriophages P1 == | ||
Revision as of 15:03, 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 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 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
