Team:Groningen/Notebook/11 August 2009
From 2009.igem.org
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m (→GVP Cluster) |
(→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
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
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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