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Team:Braunschweig/Protocols
From 2013.igem.org
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<h2><a href="#Growthcurve">Measurement of Growth Curves</a></h2> | <h2><a href="#Growthcurve">Measurement of Growth Curves</a></h2> | ||
| - | <p><p style=" margin-left:5px; margin-right:5px; margin-bottom:0px;"> | + | <p><p style=" margin-left:5px; margin-right:5px; margin-bottom:0px;">Overnight cultures in 20 mL 2xYT medium containing chloramphenicol were inoculated directly from -80°C glycerol cell stock. Cells were grown in non-baffled-flasks at 37°C and 250 rpm.<br> |
| + | The next day, main cultures in 75 mL 2xYT medium containing ampicillin were inoculated from overnight cultures to an OD520=0.5. Growth of each strain was observed with and without induction of the promoters regulation the ampicillin resistance gene. All cultures and measurements were conducted as duplicates.<br> | ||
| + | For the induction of beta-lactamase (ampR) expression by pRhl und pLas autoinducers n-buturyl-homoserinlactone and n-oxododecanoyl-homoserinlactone were added respectively to a final concentration of 10 µmol/L. Cells were grown in non-baffled flasks at 37°C and 250 rpm. <br> | ||
| + | Samples from each flask were taken at appropriate times depending on the growth phase of the cells. Optical density was measured at a wavelength of 520 nm in order to avoid/minimize absorption by the chromoproteins (see absorption spectra). Cells were left to grow until the stationary phase was reached. | ||
| + | </p> | ||
</div> | </div> | ||
Revision as of 21:52, 23 September 2013
Protocols
In this section you will find detailed protocols of experimental procedures.
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