From 2009.igem.org
Wet
GVP Cluster
- → DONE isolate plasmids from overnight precultures
- → DONE plate on cultures for storage in fridge (amp)
- → TODO cut plasmid and GVP containing plasmid for ligation (ligation tomorrow)
- → TODO purify wanted fragments (tomorrow)
- → DONE make o.n. precultures from plate colonies
Plasmid Purification
Plasmid isolation was performed on the cultures of E.coli TOP10 containing plasmids J61002 with high, medium and low constitutive promoters 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 20μL MQ and stored in the fridge
Concentrations
Plasmid
| Conc. ng/μL
| 260/280
| 260/230
| -20 box (michael
| Restriction Control
|
pJ61002-BBa_J23100 (high)
| 174.6
| 1.94
| 2.40
| E-8
| No
|
pJ61002-BBa_J23106 (medium)
| 149.2
| 1.95
| 2.41
| E-9
| No
|
pJ61002-BBa_J23109 (low)
| 152.8
| 1.94
| 2.38
| E-10
| No
|
Transporters
Metal Accumulation
Vectors
Dry