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Team:UC Chile2/Cyanolux Protocols
From 2012.igem.org
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== Methods for characterization of LuxBrick (K325909) == | == Methods for characterization of LuxBrick (K325909) == | ||
| + | <font face = "Calibri"> | ||
| + | * Pick a single colony from a transformed plate of K325909. Grow in LB media with chloramphenicol 30ug/mL O/N at 30°C. | ||
| - | + | * Measure OD at 600nm, then dilute to OD600=0.6 following direct C*V ratio to obtain bacterial stock for experiments. | |
| - | + | * Induce with 3mM (L)-Arabinose (final concentration). | |
| - | + | * Alicuot 600uL of bacteria and add LB media with chloramphenicol 30ug/mL + additives and inducers up to 1.5mL. This will yield a 0.24 OD600nm. | |
| - | Alicuot 600uL of bacteria and add LB media with chloramphenicol 30ug/mL + additives and inducers up to 1.5mL. This will yield a 0.24 OD600nm. | + | |
| - | Grow at 30°C for 1.5 hours (or at chosen T° condition for characterization). | + | * Grow at 30°C for 1.5 hours (or at chosen T° condition for characterization). |
| - | Alicuot 100uL into 4 wells (at least) of a 96 well falcon plate (plaque?). | + | * Alicuot 100uL into 4 wells (at least) of a 96 well falcon plate (plaque?). |
| - | Measure luminescence in a ThermoScientific Luminoskan Ascent Microplate Luminometer. | + | * Measure luminescence in a ThermoScientific Luminoskan Ascent Microplate Luminometer. |
| + | </font> | ||
</div> | </div> | ||
Revision as of 22:36, 13 September 2012
Methods for characterization of LuxBrick (K325909)
- Pick a single colony from a transformed plate of K325909. Grow in LB media with chloramphenicol 30ug/mL O/N at 30°C.
- Measure OD at 600nm, then dilute to OD600=0.6 following direct C*V ratio to obtain bacterial stock for experiments.
- Induce with 3mM (L)-Arabinose (final concentration).
- Alicuot 600uL of bacteria and add LB media with chloramphenicol 30ug/mL + additives and inducers up to 1.5mL. This will yield a 0.24 OD600nm.
- Grow at 30°C for 1.5 hours (or at chosen T° condition for characterization).
- Alicuot 100uL into 4 wells (at least) of a 96 well falcon plate (plaque?).
- Measure luminescence in a ThermoScientific Luminoskan Ascent Microplate Luminometer.
