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-	== Sequencing reactions ==
-
-	In order to verify the quality of the ligation reaction products (of [[Paris/September 18| Sep. 18]]), the following sequencing reactions are being performed
-
-	TL_L44.1+O18
-	TL_L44.1+O19
-	TL_L44.2+O18
-	TL_L44.2+O19
-	TL_L45.1+O18
-	TL_L45.1+O19
-	TL_L45.2+O18
-	TL_L45.2+O19
-	TL_L46.1+O18
-	TL_L46.1+O19
-	TL_L47.1+O18
-	TL_L47.1+O19
-	TL_L53.7+O18
-	TL_L53.7+O19
-	TL_L53.7+O26
-	TL_L54.3+O18
-	TL_L54.3+O19
-	TL_L54.3+O26
-	TL_L54.4+O18
-	TL_L54.4+O19
-	TL_L54.4+O26
-	TL_L56.3+O18
-	TL_L56.3+O19
-	TL_L56.3+O25
-	TL_L56.4+O18
-	TL_L56.4+O19
-	TL_L56.4+O25
-	TL_L57.3+O18
-	TL_L57.3+O19
-	TL_L57.3+O25
-	TL_L57.4+O18
-	TL_L57.4+O19
-	TL_L57.4+O25
-	TL_L58.5+O18
-	TL_L58.5+O19
-	TL_L58.7+O18
-	TL_L58.7+O19
-	TL_L59.6+O18
-	TL_L59.6+O19
-	TL_L59.6+O40
-	TL_L63.1+O18
-	TL_L63.1+O19
-	TL_L63.2+O18
-	TL_L63.2+O19
-	TL_L64.1+O18
-	TL_L64.1+O19
-	TL_L64.1+O25
-	TL_L64.2+O18
-	TL_L64.2+O19
-	TL_L64.2+O25
-
	== pDapA DAP-dependant repression through FACS analysis ==		== pDapA DAP-dependant repression through FACS analysis ==

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Revision as of 14:42, 3 October 2007

pDapA DAP-dependant repression through FACS analysis

PCR on Wanner Bfr constructions (L58.5 &L58.7) with the Bfr-Ftsk primers (O43 & O44)

• We used the Phusion high-fidelity DNA Polymerase (Finnzymes) :
  

- 26,5µL H2O
- 10µL 5X Buffer
- 7µL DNA template
- 2*2,5µL Primers
- 1µL dNTP
- 0,5µL Polymerase

• 2-step PCR reaction : the Tm was higher than 72°C, then Finnzymes asks for us to not use an annealing time anymore.
  

- 98°C initial denaturation : 1'30
30 steps of :
- 98°C denaturation : 30"
- 72°C elongation : 50"
and
- 72°C final elongation : 7'