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Team:UC Chile2/Cyanolux Protocols
From 2012.igem.org
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| + | == 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. | ||
Revision as of 02:46, 8 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.
