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Team:Groningen/Notebook/6 August 2009
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(New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== '''Over Night Cultures''' Over night cultures in 5mL LB-amp<sub>50</sub> or LB-amp<sub>25</sub>/-chloramph<sub>100</sub>...)
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(New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== '''Over Night Cultures''' Over night cultures in 5mL LB-amp<sub>50</sub> or LB-amp<sub>25</sub>/-chloramph<sub>100</sub>...)
Newer edit →
Revision as of 10:40, 6 August 2009
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Wet
GVP Cluster
Over Night Cultures
Over night cultures in 5mL LB-amp50 or LB-amp25/-chloramph100 medium were prepared from the following colonies:
- → E.coli TOP10 pSB1AC3-high const. promoter - GVP (amp.) (2x)
- → E.coli TOP10 pSB1AC3-med. const. promoter - GVP (amp.) (2x)
- → E.coli TOP10 pSB1AC3-low const. promoter - GVP (amp.) (2x)
- → E.coli TOP10 pSB1AC3-high const. promoter - GVP (amp./chl)
- → E.coli TOP10 pSB1AC3-med. const. promoter - GVP (amp./chl)
- → E.coli TOP10 pSB1AC3-low const. promoter - GVP (amp./chl)
- → E.coli TOP10 (oud) pSB1AC3-med. const. promoter - GVP (amp./chl)
and put in the 37°C waterbath at 200 rpm.
- → Note: The concentration of Chloramphe. antibiotics can be lower because crystalisation had occured during storage, vortexing did not dissolve the crystals completely!
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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