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Team:Groningen/Notebook/14 July 2009
From 2009.igem.org
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===GVP Cluster=== | ===GVP Cluster=== | ||
| + | '''Plasmid Purification''' | ||
| + | Plasmid isolation was performed on the cultures of GVP and Terminator containing cells with the "Sygma-Aldrich™ GenElute™ Plasmid Miniprep Kit". | ||
| + | * From each tube 5mL of culture was collected in a 2mL cup, and the cells were pelleted by centrifugation for 1 min. at max. speed and the supernatant discarded. | ||
| + | * Cells were resuspended in 200μL Resuspension Solution by up and down pipetting. | ||
| + | * To the mixture 200μL Lysis Solution was added, mixed by inverting the cup, and stored at room temperature for 5 minutes. | ||
| + | * 350μL of Neutralisation Solution was added and the tubes inverted. | ||
| + | * Cell debri was pelleted by centrifugation at full speed for 1 min. | ||
| + | * | ||
| + | |||
| + | '''Concentration of Plasmids''' | ||
| + | |||
| + | The concentration of isolated plasmid was determined with the use of a nano-drop. | ||
| + | |||
| + | ''GVP eluted in MQ'' | ||
| + | * ng/μL | ||
| + | * (260/280) | ||
| + | * (260/230) | ||
===Transporters=== | ===Transporters=== | ||
Revision as of 07:24, 14 July 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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Wet
GVP Cluster
Plasmid Purification
Plasmid isolation was performed on the cultures of GVP and Terminator containing cells with the "Sygma-Aldrich™ GenElute™ Plasmid Miniprep Kit".
- From each tube 5mL of culture was collected in a 2mL cup, and the cells were pelleted by centrifugation for 1 min. at max. speed and the supernatant discarded.
- Cells were resuspended in 200μL Resuspension Solution by up and down pipetting.
- To the mixture 200μL Lysis Solution was added, mixed by inverting the cup, and stored at room temperature for 5 minutes.
- 350μL of Neutralisation Solution was added and the tubes inverted.
- Cell debri was pelleted by centrifugation at full speed for 1 min.
Concentration of Plasmids
The concentration of isolated plasmid was determined with the use of a nano-drop.
GVP eluted in MQ
- ng/μL
- (260/280)
- (260/230)
Transporters
Dry
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