Team:Paris/September 2

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Construction of rbs-TetR-mRFP-LVA-tripart

Ligation L173

Samples used

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
  • Put in the room "Danger azote" at about 16°C at 18h00 (for overnight reaction).


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

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

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
  • Put in the room "Danger azote" at about 16°C at 18h00 (for overnight reaction).
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