From 2009.igem.org
Wet
GVP Cluster
- → DONE make glycerol stocks for pSB1AC3-BBa_J23106-BBa_I750016 and pSB1AC3-BBa_J23109-BBa_I750016 (two o.n. cultures of each are grown)
- → DONE isolate plasmid J61002-BBa_J23100, J61002-BBa_J23106, and J61002-BBa_J23109 from o.n. cultures
- → TODO make a planning for next week
Plates
Showed single colony growth on plates with pSB3K3-H/L-GVP plasmids, and were stored in the fridge for future preculture growth.
- → The plates with pSB1AC3-H-GVP plasmid showed no growth, and were stored at 37C for an additional 4 hours to be sure.
Over Night Cultures
- → All cultures showed growth, and can be used for plasmid isolation and glycerol stocks.
Plasmid Purification
Plasmid isolation was performed on the cultures of E.coli TOP10 containing plasmids BBa_J61002 with high, medium and low constitutive promoters with the "Sygma-Aldrich™ GenElute™ Plasmid Miniprep Kit".
- From each tube 8mL 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 50μL MQ and stored in the fridge
Concentration
Plasmid
| Conc. ng/μL
| 260/280
| 260/230
| -20 box (michael
| Restriction Control
|
BBa_J61002-BBa_J23100 (high+RFP)
| 288.1
| 1.90
| 2.33
| D-8
| No
|
BBa_J61002-BBa_J23106 (med.+RFP)
| 162.2
| 1.93
| 2.31
| D-9
| No
|
BBa_J61002-BBa_J23109 (low+RFP)
| 114.2
| 1.93
| 2.28
| D-10
| No
|
Transporters
Metal Accumulation
Vectors
Dry