Team:Paris/August 22

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Contents

Analysis of the transformant of FlhDC+promotor

KR000219.jpg
  • Analysis of the plasmids MP160.1 and MP160.2 (FlhDC+promotor in pSB1A2)
  • Control: MP143 (GFP generator in pSB1A2)

Digestion

  • 2

Construction of pFlgA - GFP Generator

Aim : Construction of "pFlgA-RBS-GFP-dbl ter" (pFlgA-E0430/E0432) Part icon regulatory.pngPart icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png

Digestion

Gel Extraction

Protocol [[Image:KR00019b.jpg| thumb|Gel Extraction of D168]

Well 1 2 3 4 5 6
Sample 100 pb ladder MP143.1 no sample D168 no sample
Expected size (pb) |
Measured size (pb)

Measurement of the concentration of D168 purified

Protocol (it's same that for Miniprep)

Digestion Name Concentration (µg/mL) Ratio 260/280
D168

Ligation

Protocol

Ligation Name Vector Name Volume Vector (µL) Insert Volume Insert (µL)
L164 D168 D132
Control L164 D168 - -


Construction of pFlgA - YFP tripart (+/- LVA)

Aim : Construction of "pFlgA-RBS-YFP-dbl ter" (pFlgA-E0430/E0432) Part icon regulatory.pngPart icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png

Transformation of the ligations we did yesterday

Protocol

Ligation Name Description Antibio
L160 D166(FV) + D132 (FI) Amp
Control L160 D166 Amp
L161 D167(FV) + D1132 (FI) Amp
Control L161 D167 Amp

Construction for synchronization

Transformation of the ligations we did yesterday

Protocol

Ligation Name Description Antibio
L158 D110(BV) + D131 (BI) Amp
T L158 D110 Amp
L159 D125(FV)+D109(FI) Kan
T L159 D125 Kan

Ligation

Protocol

Ligation Name Description Vector Name Volume vector (µL) Insert Name Volume insert(µL)
L162 rbs-lasI + gfp generator
Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png 		D107 (BV) 10 D163 (BI) 3.46
T L162 autoligation control D107 (BV) 10
L163 rbs-TetR + gfp generator
Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png 		D110 (BV) 2.5 D163 (BI) 3.44
T L163 autoligation control D110 (BV) 2.5

Transformation

Protocol

Ligation Name Description Antibio
L162 D107(BV) + D163 (BI) Amp
T L162 D107 Amp
L163 D110(BV)+D163(BI) Amp
T L163 D110 Amp
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