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The test simulates the behavior of the waste by contacting it with an acidic media during 18 h with continuous agitation.
The suspensions were then heated at 90°C for 1h with continuous agitation (200 rotations/min).
The resulting slurry was aged for 18 h with continuous agitation.
The broth was fermented at 32 °C for 72 h in an automatic incubator with continuous agitation (150 rpm).
Fresh medium was inoculated in test tubes with the starter culture and grown at 35.5°C with continuous agitation at 30 rpm.
Bacteria were regularly cultured in LB broth at 37°C with continuous agitation.
Membranes were hybridized with cDNA probes overnight at 60°C with continuous agitation at 5 10 r.p.m and then washed as recommended by the manufacturer.
E. coli harboring pBAD or pBAD/btuE plasmids (see below) were grown in LB containing ampicillin (100 µg ml−1) at 37°C with continuous agitation.
To ensure full equilibration of minerals and the specific chelator, pre-washing was maintained in EDTA-2Na·2H2O for 2.5 h with agitation during the first 30 min [42], in 2,2'-bipyridyl for 2 h with continuous agitation, and in deionized H2O for 48 h without agitation.
Hybridization was performed over-night at 60°C with continuous agitation.
One ml of the supernatant was incubated overnight with first antibody at 4°C with continuous agitation.
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