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Construction of rbs-TetR-mRFP-LVA-tripart (L173)
![Part icon rbs.png](/wiki/images/f/fe/Part_icon_rbs.png) ![Icon coding.png](/wiki/images/b/bf/Icon_coding.png) ![Part icon reporter.png](/wiki/images/a/a3/Part_icon_reporter.png) ![Part icon terminator.png](/wiki/images/1/19/Part_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.png](/wiki/images/f/fe/Part_icon_rbs.png) ![Icon coding.png](/wiki/images/b/bf/Icon_coding.png) ![Part icon reporter.png](/wiki/images/a/a3/Part_icon_reporter.png) ![Part icon terminator.png](/wiki/images/1/19/Part_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.
|