Team:Groningen/Notebook/11 August 2009
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(Difference between revisions)
m (New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== :→ {{todo}} isolate plasmids from overnight precultures :→ {{todo}} plate on cultures for storage in fridge (amp) :...) |
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:→ {{todo}} purify wanted fragments (tomorrow) | :→ {{todo}} purify wanted fragments (tomorrow) | ||
:→ {{todo}} make o.n. precultures from plate colonies | :→ {{todo}} make o.n. precultures from plate colonies | ||
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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: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". | ||
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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 06:57, 11 August 2009
Wet
GVP Cluster
- → TODO isolate plasmids from overnight precultures
- → TODO plate on cultures for storage in fridge (amp)
- → TODO cut plasmid and GVP containing plasmid for ligation (ligation tomorrow)
- → TODO purify wanted fragments (tomorrow)
- → TODO 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
Transporters
Metal Accumulation
Vectors
Dry
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