From 2008.igem.org
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- | *Put in the room "Danger azote" at about 16°C at 18h00 (for overnight reaction). | + | * overnight reaction at about 16°C. |
Revision as of 20:27, 3 September 2008
← Yesterday ↓ Calendar ↑Tomorrow →
Construction of rbs-TetR-mRFP-LVA-tripart (L173)
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)
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
|
- overnight reaction at about 16°C.
|