Team:Groningen/Notebook/25 September 2009
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(Difference between revisions)
(New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== ===Transporters=== ===Metal Accumulation=== *'''Harvest ''E.coli'' cells and As(III) uptake experiment''' **Take OD sa...) |
(→Metal Accumulation) |
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**Wash 2x with TB74S buffer. | **Wash 2x with TB74S buffer. | ||
**Resuspend upto OD=10. | **Resuspend upto OD=10. | ||
- | **Dry 1ml sample for 2hrs @ 65°C. | + | **Dry 1ml sample for 2hrs @ 65°C. |
**Do uptake experiments with the different samples (1gr cells @ 22mg/ml concentration) | **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). | ***At As(III) concentrations for 1hr: 0, 10, 20, 50 and 100uM As(III). | ||
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**Spin down, wash with TB74S and resuspend in 10ml ddH2O. | **Spin down, wash with TB74S and resuspend in 10ml ddH2O. | ||
**Dry for 2days @ 65°C. | **Dry for 2days @ 65°C. | ||
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===Vectors=== | ===Vectors=== |
Revision as of 20:10, 30 September 2009
Wet
GVP Cluster
Transporters
Metal Accumulation
- Harvest E.coli cells and As(III) uptake experiment
- Take OD sample to deterimine OD600.
- Harvest E.coli WT (pSB1A2 + pLac), E.coli + pArsR-RFP and E.coli + pLac-fMT by
spinning down @ 6000rpm for 20min in Sorvall centrifuge.
- Wash 2x with TB74S buffer.
- Resuspend upto OD=10.
- Dry 1ml sample for 2hrs @ 65°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°C.
Vectors
Dry
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