Seventy-seven dusky kob fingerlings (mean body mass 9.14 ± 0.30 g) were distributed into each of 20 experimental tanks and offered diets at 2.8% body mass.
For the bacterial challenge test, ten apparently healthy prawns of similar size (1.2 1.5 g) were distributed into each triplicate tank and fed their respective experimental diet twice daily (8 30 and 16 30 h) for 1 week.
For siRNA transfections, a master mix of Serum- and Phenol Red-free DMEM (7 µL/transfection), HiPerFect reagent (1 µL/transfection, Qiagen) and 2 µL of a 10 µM siRNA solution (2 pmols/transfection) was distributed into each well of a 384-well plate and incubated at room temperature for 20 40 minutes.
Fresh media was distributed into each well followed by a 15 minute incubation at 37°C and 5% CO2.
For overexpression of FLAG-tagged constructs, plasmid (2 µg) was distributed into each well along with FuGENE HD (4 μl, Roche; Switzerland).
Glass balls with 2 mm in diameter (Hecht) were distributed into each well of the 96-well plate to increase agitation.
Then, 100 µL of yeast transformation mix was distributed into each well of a 96-well half skirt PCR plate (MPX-96M2; Phenix Research Products), after which 10 µL of MoBY-ORF plasmid was added and repeatedly mixed using the BioMek FX Laboratory Automation Workstation/Biomek Software.
Target cells were resuspended in medium at 1 × 105 cells/ml, and 100 μl of 105 cells/ml cell suspension were distributed into each well of 96-well plates (Costar) and allowed to adhere for 24 hours.
MCF-7 breast cancer cells were resuspended in medium at 1 × 105 cells/ml, and 100 μl of 105 cells/ml cell suspension were distributed into each well of 96-well plates (Costar, Cambridge, MA, USA) and allowed to adhere for 24 hours.
Let the available bandwidth be distributed into U subchannels, where each subchannel contains K/U subcarriers.
A total of 43 ml of blood will be collected from each volunteer to be distributed into 5 vacutainers.
