Team:Paris/September 2

From 2008.igem.org

(Difference between revisions)
(→Ligation L172)
(→Ligation L173)
 
Line 6: Line 6:
=='''Ligation L173'''==  
=='''Ligation L173'''==  
{|border="1" style="text-align: center"
{|border="1" style="text-align: center"
 +
|'''Digestion n°'''
|'''Name'''
|'''Name'''
-
|'''Digestion number'''
 
|'''Digested by'''
|'''Digested by'''
|'''DNA concentration'''
|'''DNA concentration'''
|-
|-
-
|rbs-TetR
 
|D112
|D112
 +
|rbs-TetR
|EcoRI & SpeI (insert)
|EcoRI & SpeI (insert)
|~20 µg/mL
|~20 µg/mL
|-
|-
-
|mRFP-LVA-tripart
 
|D187
|D187
 +
|mRFP-LVA-tripart
|EcoRI & XbaI (vector)
|EcoRI & XbaI (vector)
|16 µg/mL
|16 µg/mL

Latest revision as of 19:24, 9 September 2008

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Contents

Construction of rbs-TetR-mRFP-LVA-tripart (L173)

Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png

Ligation L173

Digestion n° Name Digested by DNA concentration
D112 rbs-TetR EcoRI & SpeI (insert) ~20 µg/mL
D187 mRFP-LVA-tripart EcoRI & XbaI (vector) 16 µg/mL


Reaction mixtures

Control Sample
3:1 (insert:vector) ratio 4:1 (insert:vector) ratio
D112 (insert) 0 µL 1,4 µL 1,9 µL
D187 (vector) 2,5 µL 2,5 µL 2,5 µL
T4 DNA ligase 10X buffer 2 µL 2 µL 2 µL
T4 DNA ligase 1 µL 1 µL 1 µL
water 14,5 µL 13,1 µL 12,6 µL
  • overnight reaction at about 16°C.


Construction of rbs-FlhDC-mRFP-LVA-tripart (L172)

Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png

Ligation L172

Digestion n° Name Digested by DNA concentration
D186 rbs-FlhDC EcoRI & SpeI (insert) ~24 µg/mL
D187 mRFP-LVA-tripart EcoRI & XbaI (vector) 16 µg/mL


Reaction mixtures (3:1 (insert:vector) ratio)

Control Sample
D186 (insert) 0 µL 1 µL
D187 (vector) 2,5 µL 2,5 µL
T4 DNA ligase 10X buffer 2 µL 2 µL
T4 DNA ligase 1 µL 1 µL
water 14,5 µL 13,5 µL
  • overnight reaction at about 16°C.
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