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Team:Groningen/Notebook/7 August 2009
From 2009.igem.org
(Difference between revisions)
(New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== :→ {{todo}} isolate plasmids from overnight precultures :→ {{todo}} perform a restriction on isolated plasmid (shoul...) |
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:→ {{todo}} cut pSB3K3 plasmid and GVP containing plasmid for ligation | :→ {{todo}} cut pSB3K3 plasmid and GVP containing plasmid for ligation | ||
:→ {{todo}} purify wanted fragments | :→ {{todo}} purify wanted fragments | ||
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| + | [[Image:Plasmid Miniprep Kit Flow Protocol.gif|thumb|150px| www.sigmaaldrich.com]] | ||
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| + | '''Plasmid Purification''' | ||
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| + | Plasmid isolation was performed on the cultures of E.coli TOP10 containing plasmids [http://partsregistry.org/wiki/index.php?title=Part:pSB3K3 pSB3K3] with high, medium and low constitutive promoters and [http://partsregistry.org/wiki/index.php?title=Part:BBa_I750016 BBa_I750016] with the "Sygma-Aldrich™ [http://www.sigmaaldrich.com/life-science/molecular-biology/dna-and-rna-purification/plasmid-miniprep-kit.html GenElute™] Plasmid Miniprep Kit". | ||
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| + | * 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 | ||
===Transporters=== | ===Transporters=== | ||
Revision as of 07:16, 7 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
- → TODO isolate plasmids from overnight precultures
- → TODO perform a restriction on isolated plasmid (should result in 6000 and 3000bp fragments)
- → TODO plate on cultures for pure colony over the weekend
- → TODO cut pSB3K3 plasmid and GVP containing plasmid for ligation
- → TODO purify wanted fragments
Plasmid Purification
Plasmid isolation was performed on the cultures of E.coli TOP10 containing plasmids [http://partsregistry.org/wiki/index.php?title=Part:pSB3K3 pSB3K3] with high, medium and low constitutive promoters and [http://partsregistry.org/wiki/index.php?title=Part:BBa_I750016 BBa_I750016] 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
Transporters
Metal Accumulation
Vectors
Dry
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