Team:Groningen/Notebook/25 September 2009

Metal Accumulation

 * Harvest E.coli cells and As(III) uptake experiment

spinning down @ 6000rpm for 20min in Sorvall centrifuge.
 * Take OD sample to deterimine OD600.
 * Harvest E.coli WT (pSB1A2 + pLac), E.coli + pArsR-RFP and E.coli + pLac-fMT by
 * Wash 2x with TB74S buffer.
 * Resuspend upto OD=10.
 * Dry 1ml sample for 2hrs @ 65&deg;C.
 * Do uptake experiments with the different samples (1gr cells @ 22mg/ml concentration)
 * At As(III) concentrations for 1hr: 0, 10, 20, 50 and 100uM As(III).
 * And at time points with 10uM As(III) at: 0, 10, 20, 40 and (60)min.
 * For fMT only the concentration samples were taken!
 * Spin down, wash with TB74S and resuspend in 10ml ddH2O.
 * Dry for 2days @ 65&deg;C.


 * The determined dry weight was 5x lower than expected, so the sample which should have contained at least 1gr dry cells only contained ~100-200mg dry cells.