Team:Paris/August 22

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(Difference between revisions)
(→Construction of pFlgA - YFP tripart (+/- LVA))
(→Construction of pFlgA - YFP tripart (+/- LVA))
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=Analysis of the transformant of FlhDC+promotor=
=Analysis of the transformant of FlhDC+promotor=
 +
 +
='''Construction of pFlgA - GFP Generator'''=
 +
 +
Aim : Construction of ''' "pFlgA-RBS-GFP-dbl ter" (pFlgA-E0430/E0432)''' [[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 +
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==Digestion==
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==='''Gel Extraction'''===
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[[Team:Paris/Notebook/Protocols#Migration after extraction |Protocol]]
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[[Image:KR00019b.jpg| thumb|Gel Extraction of D166-D167]
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{|border="1" style="text-align: center"
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|'''Well'''
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|1
 +
|2
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|3
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|4
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|5
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|6
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|7
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|8
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|-
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|'''Sample'''
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|100 pb ladder
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|MP143.1
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|no sample
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|colspan="2"|D168
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|colspan="3"|no sample
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|-
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|'''Expected size (pb)'''
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|
 +
|
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|
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|colspan="2"| |
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|-
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|'''Measured size (pb)'''
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|
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|style="background: #cbff7B"|
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|
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|colspan="2"| style="background: #cbff7B"|
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|
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|}
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==='''Measurement of the concentration of D168 purified'''===
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[[Team:Paris/Notebook/Protocols#Concentration_of_the_Miniprep | Protocol (it's same that for Miniprep)]]
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 +
{|border="1" style="text-align: center"
 +
|'''Digestion Name'''
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|'''Concentration (µg/mL)'''
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|'''Ratio 260/280'''
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|-
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|D168
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|
 +
|
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|}
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==Ligation==
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[[Team:Paris/Notebook/Protocols#Ligation |Protocol]]
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 +
{|border="1" style="text-align: center"
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|'''Ligation Name'''
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|'''Vector Name'''
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|'''Volume Vector (µL)'''
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|'''Insert'''
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|'''Volume Insert (µL)'''
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|-
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|L164
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|D168
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|
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|D132
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|
 +
|-
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|Control L164
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|D168
 +
|
 +
| -
 +
| -
 +
|}
 +
='''Construction of pFlgA - YFP tripart (+/- LVA)'''=
='''Construction of pFlgA - YFP tripart (+/- LVA)'''=

Revision as of 16:00, 22 August 2008

← Yesterday

↓ Calendar ↑

Tomorrow →

Contents

Analysis of the transformant of FlhDC+promotor

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 D166-D167]

Well 1 2 3 4 5 6 7 8
Sample 100 pb ladder MP143.1 no sample D168 no sample
Expected size (pb) |
Measured size (pb) style="background: #cbff7B"|


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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