Team:Groningen/Notebook/16 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

Plates


 * → The plates showed a low amount of colonies, and no colonies in the case of the negative plates.


 * → The two plates were stored in the fridge for further use.

Cultures

The overnight cultures with LB-amp100 medium of colonies E.coli TOP10 with pLacI-GVP (no.1 and 2), pZntR-GVP (glycerol), pCueO-GVP (glycerol), GlpF (no.1 and 2), and pLacI (pSB1AC3) 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 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

The plasmids from the o.n. precultures of pArsR-GVP and pSB2K3 (earlier last week) were cut with PstI and EcoRI to cut out the entire part between the pre- and suffix. The plasmids with PstI were cut with PstI for control.




 * → From left to right: 1kB ladder, three samples of SJ, pCueO-GVP (pSB2K3), pZntR-GVP (pSB2K3), pLacI-GVP (pSB1A2, no.1), pLacI-GVP (pSB1A2, no.2), GlpF (pSB1AC3), pLacI (pSB1AC3)


 * → The additional two lanes are a darker image of the GlpF (pSB1AC3), pLacI (pSB1AC3) lanes, to check for GlpF and pLacI insert.