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One hundred microliters of a 5 × 10/ml O4GIP ESC solution in 1.5× concentrated 2i in N2B27 was dispensed in each well using a semi-automated cell dispenser (Genetix Cell Dispense).
Two hundred microliter of the cell suspension were dispensed in each well of a 96-well flat bottom microplate with the addition of Doxy and BCM when needed.
Count cells and dilute them in fresh medium so that the desired number of cells is in 30-40 µL of medium that can be dispensed in each well.
One drop of antifoaming agent, antifoam B emulsion, was dispensed in each flask after initial foaming was observed during aeration.
Aliquot (50 μl) of either ROS-BSA conjugate or BSA protein solution (5 μg/ml in PBS) was dispensed in each well of the microwell plate.
A 50 μl of anti-ROS antibody sample (rabbit serum or purified IgG) was dispensed in each well and plate was incubated for 1.5 h at 37°C, then the plates were washed with PBS-T, and 50 μl of HRP-IgG (1/5,000 in PBS) was added to each well.
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Between songs, Mr. Spooner dispensed in-crowd banter like: "Was it intentional?
Randomization of the doses was pre-determined and built into the packaging, so that each numbered box of medication was dispensed in sequence to each enrolled patient.
Leucoperm A and Leucoperm B (Serotec, UK) were added at RT for 15 min and cells were dispensed in four tubes each containing 1 × 10 cells/100 μL.
Firstly, the cells were processed as described in the supplementary information, dispensed in 96-well plates, each one containing 5 × 104 cells, and kept in culture for 24 h.
Drugs were dispensed in patient-wise boxes each containing blister packs for daily and weekly administration in IP and CP respectively.
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