Paris/October 3
From 2007.igem.org
(→Culture for miniprep of S49 having the pKD46 plamid) |
(→Culture for miniprep of a strain (S51) having the plasmid pY-2P-IntC) |
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== Culture for miniprep of a strain (S51) having the plasmid pY-2P-IntC == | == Culture for miniprep of a strain (S51) having the plasmid pY-2P-IntC == | ||
+ | |||
+ | this plasmid will help us to integrate the [pTet>>lox71-GFPtripart-lox66-mRFP] construction into IntC site on the bacteria chromosome. This construction will permit to undergo some recombination rate experiments on the future strain. | ||
== The wanner PCR didn't give the right size, exploration for explanation == | == The wanner PCR didn't give the right size, exploration for explanation == |
Revision as of 17:29, 3 October 2007
Contents |
Sequencing reactions
In order to verify the quality of the ligation reaction products (of 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_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
Transformation of W121 and FR781 with pKD46 didn't work
We got no clones. After reflexion, it appeared that the miniprep of pKD46 i did came from a 37°C culture, and pKD46 is thermosensible, the explanation is just that i transformed bacteria with no plasmid.
i launched a new culture of S49 at 30°C this time.
Culture for miniprep of S49 having the pKD46 plamid
with 5mL of Amp-LB at 30°C.
Culture for miniprep of a strain (S51) having the plasmid pY-2P-IntC
this plasmid will help us to integrate the [pTet>>lox71-GFPtripart-lox66-mRFP] construction into IntC site on the bacteria chromosome. This construction will permit to undergo some recombination rate experiments on the future strain.