Team:Groningen/Notebook/11 August 2009
From 2009.igem.org
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(→GVP Cluster) |
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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. | * 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 | * Plasmids were eluted with 20μL MQ and stored in the fridge | ||
+ | |||
+ | '''Concentrations''' | ||
+ | |||
+ | {|cellpadding="2" cellspacing="1" border="4" | ||
+ | |'''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=== | ===Transporters=== |
Revision as of 08:21, 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
- → 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 [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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