Each well contained two layers of food medium with the top layer containing luciferin.
The medium at the top layer was changed on every sixth day.
Samples for microbial characterization were collected from the filter medium (top and bottom layers) of the wetland, water influent and effluent at the end of Periods I (Jun Oct) and II (Nov Jan).
On day 4, the top layer medium was changed with fresh medium prepared as previously mentioned.
The top layer medium was prepared by adding the following to M199 basal medium: 20% of heat inactivated fetal bovine serum (HIFBS) (Gibco, Invitrogen, USA), 1% L-glutamine (Lonza, Belgium), 0.1% aminocaproic acid (Sigma, USA), 1% amphotericin B (Lonza, Belgium) and 0.6% gentamicin (Lonza, Belgium).
Plant extracts were added to the top layer medium at concentration of 100 μg/mL.
The protoplasts were collected by centrifugation at 180g for 2 min at room temperature, and suspended in 8 ml of top layer protoplast regeneration medium (BCD medium supplemented with 5 mM ammonium tartrate, 10 mM CaCl2, 0.8% agar and 8% mannitol) preheated at 45 °C.
The road pavement can be modeled as a beam, a plate, or the top layer of a layered soil medium.
For example, in a two-layer medium, the thickness of the top layer is generally required in order to derive reasonably precise estimates of the optical properties [ 9].
In six-well plates, each well contained a bottom layer of 1% agarose, a middle layer of 0.5% agarose which included 7.5 × 10 cells, and a top layer of medium.
