Team:Groningen/Notebook/17 September 2009

GVP Cluster
Planning


 * → work out the wiki page for GVP
 * → ✅ make a doodle for presentation planning (1-19 oct.)
 * → media attention
 * → ✅ place an ethics survey link on twitter


 * → clone pArsR-GVP into pSB2K3
 * → clone repeat out of GVP cluster
 * → make glycerol stocks of constructs
 * → enter info on part registry

Cultures

The overnight cultures with LB-amp100 medium of colonies E.coli TOP10 with pNL29 (3x), and pArsR-GVP (1-3) all showed expected growth of bacteria.



Plasmid isolation

Plasmid isolation was performed on the cultures of E.coli TOP10 containing the above mentioned plasmids with the "Sygma-Aldrich™ GenElute™ Plasmid Miniprep Kit".


 * From each tube 4mL of culture (8mL for pNL29) was collected in a 2.0mL cup (tubes from pArsR-GVP, GVP and pNL29 were combined), and the cells were pelleted by centrifugation for 1 min. at max. speed and the supernatant discarded.
 * Plasmids were eluted with 30μL MQ and stored in the fridge

Concentrations

Restriction for Control and Assembly

The plasmids from the o.n. precultures of pArsR-GVP and pNL29 were cut with PstI/EcoRI and MvaI/XhoI to cut out the entire part between the pre- and suffix and wanted fragment.




 * → From left to right: 1kB ladder, pNL29 (orange dye), pNL29 (blue dye), pArsR-GVP no.1, pArsR-GVP no.2, pArsR-GVP no.3