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September 14, 2007 saw the very first Friday Talking Points column ever (although the name and the column format wouldn't solidify for a few months).
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In our microarray experiment, swarm center and swarm tip bacteria were compared to a control colony grown on the same media solidified for a slightly longer drying period, thus avoiding gene expression differences specific to the surface vs. broth lifestyles.
The coverslips were removed and 75 μl of this second layer was quickly pipetted onto a slide and allowed to solidify for 5 min under a fresh coverslip.
The resulting solution was mixed immediately and dispensed into 4-well plates (Nunc) and allowed to solidify for 1 hour inside a 37°C, 5% CO2 incubator.
Growth factor-reduced Matrigel™ (BD Biosciences) was combined in a 1 1 ratio with differentiation medium; 100 μL was added to each well of an eight-well glass slide chamber (Lab-Tek; Thermo Scientific) and allowed to solidify for 2 hours in a 37°C incubator.
If the working area is very dry, let the plates solidify for 2 hours then cover with a tray to maintain humidity.
The mixture was then poured onto a layer of bottom agar and allowed to solidify for 5 minutes.
The chambers were placed in a cell culture incubator to allow the basement membrane to solidify for at least 20 minutes.
Allow the gel to solidify for at least 1-2 hours.
Gels were allowed to solidify for 30 min at 37°C.
Allow the glue to solidify for 20 min.
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