Team:Paris/September 2

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Revision as of 20:30, 3 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

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

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