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Team:Groningen/Notebook/11 August 2009
From 2009.igem.org
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:→ {{done}} isolate plasmids from overnight precultures | :→ {{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}} cut plasmid and GVP containing plasmid for ligation (ligation tomorrow) | ||
:→ {{todo}} purify wanted fragments (tomorrow) | :→ {{todo}} purify wanted fragments (tomorrow) | ||
| - | :→ {{ | + | :→ {{done}} make o.n. precultures from plate colonies |
[[Image:Plasmid Miniprep Kit Flow Protocol.gif|thumb|150px| www.sigmaaldrich.com]] | [[Image:Plasmid Miniprep Kit Flow Protocol.gif|thumb|150px| www.sigmaaldrich.com]] | ||
Revision as of 09:20, 11 August 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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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 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J61002 J61002] with [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 high], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 medium] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 low] constitutive promoters with the "Sygma-Aldrich™ [http://www.sigmaaldrich.com/life-science/molecular-biology/dna-and-rna-purification/plasmid-miniprep-kit.html 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
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