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Revision as of 11:00, 12 July 2007 (view source) Nicolas C. (Talk | contribs) ← Older edit		Latest revision as of 17:45, 7 October 2007 (view source) Nicolas C. (Talk | contribs)
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-	David B:	+	[[Paris/July 11|yesterday]] -- [[Paris/July 13|tomorrow]] <br>
-		+
	==Preparation of a stock of phage on w121:==		==Preparation of a stock of phage on w121:==
-	* Filtration of the Cacl2 and MgSO4 stocks	+	See [[Paris/PROTOCOLS#Preparation_of_the_P1_stock_on_the_w121_strain.|Protocols]]
-	* 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 1ml of the ON culture of W121	+
-	* Incubate 2h	+
-	* Add 200µl Chloroform	+
-	* Vortex, centrifuge (15min @ 3000rpm)	+
-	* Recover the supernatant = Stock keep at 4°C	+
-	[[User:Nicolas C.|Nicolas C.]]
	==Titration of bacteriophages P1 ==		==Titration of bacteriophages P1 ==
		+	[[User:Nicolas C.|Nicolas C.]]<br>
	We measure the strength of former P1 preparation :		We measure the strength of former P1 preparation :
	* The stock prepared the 3.7.7		* The stock prepared the 3.7.7
	* The stock prepared the 8.7.7		* The stock prepared the 8.7.7
-	See protocols for details.	+	* The stock prepared today (12.7.7)
		+	See [[Paris/July_13|results]].<br>
		+	See [[Paris/PROTOCOLS#Titration of bacteriophages|protocols]] for details.
		+
		+	== Culture of FtsZ TS ==
		+	[[User:david.bikard|David B]]<br>
		+	We want to isolate a good clone of the strain FstZ TS <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 <br>
		+	If one clone do not grow at 42, we'll use it to do the transduction. <br>
		+	See [[Paris/July_13#Test of Ftsz TS strain| 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. Without Nile red, the colonies show no fluorescence.
		+	<br>
		+	[[Image:Paris_Acineto_Colonies.jpg|500px|center]]
		+
		+	== E.coli pKS::DGAT on LB oleate medium ==
		+
		+	We incubated E.coli pKS::DGAT overnight on different LB medium (see [[Paris/July_11|July 11]]).
		+
		+	We did not observe any colony. Probably because spread plates with liquid culture kept in the freezer for one week was not a good idea...
		+
		+	On minimum medium, it did not grow too because what we thought to be M9 medium was only agar...

---

Latest revision as of 17:45, 7 October 2007

yesterday -- tomorrow

Contents 1 Preparation of a stock of phage on w121: 2 Titration of bacteriophages P1 3 Culture of FtsZ TS 4 Acinetobacter microscopy 5 E.coli pKS::DGAT on LB oleate medium

Preparation of a stock of phage on w121:

See Protocols

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. Without Nile red, the colonies show no fluorescence.

E.coli pKS::DGAT on LB oleate medium

We incubated E.coli pKS::DGAT overnight on different LB medium (see July 11).

We did not observe any colony. Probably because spread plates with liquid culture kept in the freezer for one week was not a good idea...

On minimum medium, it did not grow too because what we thought to be M9 medium was only agar...