From 2009.igem.org
Wet
GVP Cluster
BBa_F2620
O.n. precultures
- → All four o.n. precultures showed bacterial growth, and could be used to isolate plasmid. The four are registry vector pSB1A2 with pLacI/pBad-araC and GVP. The isolated plasmid is used for transformation into pSB2K3 and sent for sequencing.
Plasmid Purification
Plasmid isolation was performed on the cultures of E.coli TOP10 containing the above mentioned plasmids with the "Sygma-Aldrich™ GenElute™ Plasmid Miniprep Kit".
- From each tube 4mL of culture was collected in a 2.0mL cup, and the cells were pelleted by centrifugation for 1 min. at max. speed and the supernatant discarded.
- Plasmids were eluted with 30μL MQ and stored in the fridge
Concentrations
Plasmid
| Conc. ng/μL
| 260/280
| 260/230
| -20 box (michael
| Restriction Control
|
pSB1A2 pLacI-GVP no.1
| 405.3
| 1.85
| 2.36
| D-6
| Yes (EcoRI/PstI)
|
pSB1A2 pLacI-GVP no.2
| 34.0
| 2.01
| 2.20
| D-7
| Yes (EcoRI/PstI)
|
pSB1A2 pBad/araC-GVP no.1
| 27.5
| 2.09
| 2.16
| D-8
| Yes (EcoRI/PstI)
|
pSB1A2 pBad/araC-GVP no.2
| 33.3
| 1.96
| 2.06
| D-9
| Yes (EcoRI/PstI)
|
Transporters
Metal Accumulation
Vectors
Dry