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Team:Groningen/fluorimetry
From 2011.igem.org
Fluorimetry
Fluorimetry protocol
M9 minimal media
The OD600 and fluorescence of a great number of samples can be measured the same time with the fluorimeter.
We used it for measuring PBADaraC-cI-LVA+PcI-GFP with different concentrations of arabinose as an induction
signal
Fluorimetry protocol
Inoculate 10ml M9 medium+ antibiotics + 1ml 2% casamino acids and grow the cultures overnight at 37°C.
Grow a logfase to an OD600 of 0.4.
Dilute in a way that the OD600 is decreased to a value of 0.004/
Add 300μl of the diluted culture into a well.
The fluorimeter will measure the OD600+ fluorescence every 15 minutes.
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M9 minimal media
Component Concentrations
1x M9 salts
2mM MgSO4
0.1mM CaCl2
0.4% carbon source (e.g. glycerol, glucose, etc.)
In sterile H2O
Protocol
For 1L of media
500ml 2xM9 salts ∗
20ml 0.1M MgSO4 ∗
200μl 0.5M CaCl2 ∗
469.8ml sterile deionized H2O ∗ (if carbon source to be added)
479.8ml sterile deionized H2O ∗ (if no carbon source to be added, useful as a control for sugar metabolism)
Combine above solutions using sterile technique. (May notice some precipitation during preparation but precipitate
should go back into solution once volume is brought up to 1l with sterile H2O.)
Add antibiotic as appropriate and store at 4°C
∗ Can be obtained from the media room
http://openwetware.org/wiki/Endy:M9_medium/minimal
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