Revision as of 01:32, 22 September 2013

Notebook : August 30

check the size by Gel electrophoresis for PCR clonies on Gibson assembly and ligation promoter fnr(repressor) plus RBS_AmilCP+Term_PSB1C3 .
Do ligation for For promoter fnr(repressor)+ RBS_LacZ+Term_PSB1C3 , promoter fnr(activator)nirK + RBS_LacZ+Term_PSB3K3 and promoter fnr(repressor) + RBS_LacZ+Term_PSB3K3 and Transformation and

incubation.

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Well 1 : 6µL DNA Ladder
Well AA 1-8 : Clonies from Gibson FNR_part1, FNR part1 and plasmid PSB1C3
Well AB 1-8 : Clonies from Gibson FNR_part1, FNR part1 and plasmid PSB1C3 Concentrate3
Well AC 1-8 :Clonies from Gibson RBS_BphR2_part1, BphR2_part2 and plasmid PSB1C3
Well AD 1-8 :Clonies from Gibson RBS_FNR part1, FNR_part2 and plasmid PSB1C3
Well AE 1-8 :Clonies from Gibson RBS_FNR part1, FNR_part2 and plasmid PSB1C3 Concentrate
Well AF 1-8 :Clonies from Gibson RBS_BphR2_part1, BphR2_part2 and plasmid PSB1C3 Concentrate

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Ligation

For promoter fnr(repressor)+ RBS_LacZ+Term_PSB1C3 :

Transformation theligation and spread in LB plates with Chlorenphenicol and Xgal. incubated at 37°C in aerobic condition over night.

For promoter fnr(activator)nirK + RBS_LacZ+Term_PSB3K3 :

Transformation theligation and spread in LB plates with k and Xgal. incubated at 37°C in anaerobic condition over night.

For promoter fnr(repressor) + RBS_LacZ+Term_PSB3K3 :

Transformation theligation and spread in LB plates with k and Xgal. incubated at 37°C in aerobic condition over night.

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@@ Line 165: / Line 165: @@
 XiaoJing
+{|
+| style="border:1px solid black;padding:5px;background-color:#DE;" |
 IT WORKS !!!
+|}
 [[File:PsPfnr3008.jpg|600px]]
 In anaerobic conditions, thank to promotor RBS-Amil CP-Term, the dectetion gene is activated and we can see a violet coloration of bacterias.
 In aerobic conditions, promotor RBS-Amil CP-Term is inactive, the dectetion gene isn't activated and we can't see any coloration of bacterias.
+===='''2 - Results of culture of Pfnr-RBS-Amil CP-Term in PSB1C3, NirB with RBS-LacZ-Term in PSB1C3 in aerobic and anaerobic conditions'''====
+XiaoJing
+{|
+| style="border:1px solid black;padding:5px;background-color:#DE;" |
+Purification of 08/22/13 didn't work. We have blue colonies for Pfnr with RBS-Amil CP-Term in PSB1C3 in aerobic and anaerobic conditions. We also have blue colonies for NirB with RBS-LacZ-Term in PSB1C3 in anaerobic conditions.
+|}
+[[File:PsPfnrcult3008.jpg|500px]]
+[[File:PsNirBcult3008.jpg|500px]]
 ===='''1 - Ligation of Pfnr with RBS-LacZ-Term in PSB1C3 '''====