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Team:Groningen/Notebook/10 September 2009
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Wet
GVP Cluster
O.n. precultures
- → All tubes for plasmid isolation (pLacI (pSB1A2)(no.1 and no.2), pSB2K3 (12J)(2x) and pSB2K3 (12L)(2x) showed growth. The pSB2K3 had less growth (and smaller pellet) than the other tubes.
Plasmid isolation
Concentration
Restriction for Assembly
The plasmids from the o.n. precultures were cut with PstI and EcoRI to cut out the entire part between the pre- and suffix.
| Plasmid | Amount μL | MQ μL | Fast digest buffer | EcoRI fast digest enzyme | XbaI fast digest enzyme | SpeI fast digest enzyme | PstI fast digest enzyme |
| pLacI no.1 (pSB1A2) | 16.0 | x | 3.0 | x | x | 1.0 | 1.0 |
| pLacI no.1 (pSB1A2) | 16.0 | x | 3.0 | x | x | 1.0 | 1.0 |
| GVP | 4.0 | 11.0 | 3.0 | x | 1.0 | x | 1.0 |
| pSB2K3 (12J) | 8.0 | 6.0 | 3.0 | 1.0 | x | x | 1.0 |
| pSB2K3 (12L) | 4.0 | 11.0 | 3.0 | 1.0 | x | x | 1.0 |
| pZntR-GVP | 12.0 | 3.0 | 3.0 | 1.0 | x | x | 1.0 |
- → From left to right: 1kB ladder, pZntR-GVP (2x), Empty Slot, pSB2K3 12J (2x), Empty Slot, pSB2K3 12L (2x)
- → From left to right: 1kB ladder, pLacI no.1 (pSB1A2)(2x), pLacI no.2 (pSB1A2)(2x), GVP (2x), Empty Slot,
Transporters
Metal Accumulation
Vectors
Dry
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