From 2009.igem.org
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 10 min.
- Column was prepared by adding 500μL Column Preparation Solution and centrifuging for 1 min. full speed
- The lysate was transfered to the column and centrifuged for 1 min. full speed
- Column was washed with both washing solutions
- Plasmids were eluted with 30μL MQ and stored in the fridge
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