Team:Paris/August 23

From 2008.igem.org

(Difference between revisions)
(→digestion)
(→Digestion)
Line 108: Line 108:
=='''Digestion'''==
=='''Digestion'''==
[[Team:Paris/Notebook/Protocols#Digestion |Protocol]]
[[Team:Paris/Notebook/Protocols#Digestion |Protocol]]
 +
 +
{| border="1" style="text-align: center"
 +
|'''Name'''
 +
|'''Template DNA'''
 +
|'''Description'''
 +
|'''Vol MP (µl)'''
 +
|'''Vol H2O (µl)'''
 +
|'''Enzymes'''
 +
|-
 +
|D107
 +
|MP105.1
 +
|rbs-LasI (BV)
 +
|14.7 (0.5µg of plasmid)
 +
|11
 +
|SpeI / PstI
 +
|-
 +
|D109
 +
|MP105.1
 +
|rbs-LasI (FI)
 +
|14.7
 +
|11
 +
|EcoRI / SpeI
 +
|-
 +
|D110
 +
|MP106
 +
|rbs-TetR (BV)
 +
|9
 +
|15.7
 +
|SpeI / PstI
 +
|-
 +
|D125
 +
|MP118.1
 +
|Double terminator (FV)
 +
|4.54
 +
|20.16
 +
|EcoRI and XbaI
 +
|-
 +
|D163
 +
|MP143
 +
|gfp generator  (BI)
 +
|6.66
 +
|18.04
 +
|XbaI and PstI
 +
|}

Revision as of 12:26, 25 August 2008

← Yesterday

↓ Calendar ↑

Tomorrow →


Contents

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

Transformation of the ligations we did yesterday

Protocol

Ligation Name Description Antibio
L164 D168 (FV) + D132 (FI) Amp
Control L164 D168 Amp

Construction for Synchronization

Results of the transformation we did yesterday

Number of colonies

Ligation name Description Antibio Number Colonies observed Fluorescence Comments
Ligations
L158 D110(BV) - D131(BI)
rbs-TetR - gfp tripart 		Amp - - to do again
L159 D125(FV) - D109(FI)
rbs-LasI - double terminator 		Kan - - to do again
L162 D107(BV) - D163(BI)
rbs-LasI - gfp generator 		Amp - - to do again
L163 D110(BV) - D163(BI)
rbs-TetR - gfp generator 		Amp - - to do again
Controls
TL158 D110(BV) Amp - - to do again
TL159 D125(FV) Kan - - to do again
TL162 D107(BV) Amp - - to do again
TL163 D110(BV) Amp - - to do again
Positive Control pUC19 Amp 358 (efficiency 3.5.10^7) No OK 
=> We decided to do again the digestions of the ligation's reactions

Digestion

Protocol

Name Template DNA Description Vol MP (µl) Vol H2O (µl) Enzymes
D107 MP105.1 rbs-LasI (BV) 14.7 (0.5µg of plasmid) 11 SpeI / PstI
D109 MP105.1 rbs-LasI (FI) 14.7 11 EcoRI / SpeI
D110 MP106 rbs-TetR (BV) 9 15.7 SpeI / PstI
D125 MP118.1 Double terminator (FV) 4.54 20.16 EcoRI and XbaI
D163 MP143 gfp generator (BI) 6.66 18.04 XbaI and PstI
Retrieved from "http://2008.igem.org/Team:Paris/August_23"