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The upper tray plate of an Omni tray (Nunc) was cut in a rectangular shape and was fixed to a thin glass cover plate (Matsunami, No. 1, 73 mm × 118 mm, 0.1 mm thickness) instead of a plastic board.
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Based on percent survival, a total of 1.1 × 10 cells were present on the 260 tray plates, 3.3 × 10 cells on the 40 tray plates from the 2-h irradiation, 5.8 × 10 cells on the 80 tray plates from the 4-h irradiation, and 1.8 × 10 cells on the 140 tray plates from the 6-h irradiation.
After 4 h, 5-mL samples were again taken from each trough plate and 10-μL aliquots spread onto 40 tray plates for a total of 80 tray plates from both trough plates.
After 6 h, the remaining 10 mL was taken from each trough plate and 10-μL aliquots from the trough plate from Fernbach flask 1 were spread onto 80 tray plates and 10-μL aliquots from the trough plate from Fernbach flask 2 were spread onto 60 tray plates for a total of 140 tray plates from both trough plates.
A total of 1.1 × 10 cells were screened, 3.3 × 10 cells on the 40 tray plates from the 2-h irradiation, 5.8 × 10 cells on the 80 tray plates from the 4-h irradiation, and 1.8 × 10 cells on the 140 tray plates from the 6-h irradiation.
After incubation of the tray plates for 48 h, they were screened visually to select the colonies that produced the most intense blue color with the indicator.
The tray plates were screened visually to select colonies with the most intense blue color indicating high levels of ammonia production.
After 2 h of irradiation, 5-mL samples were taken from each trough plate and 10-μL aliquots spread onto twenty 128 × 96 mm Omni Tray plates (Thermo Fisher Scientific, Waltham, MA) for a total of 40 tray plates from both trough plates.
Samples of the surviving cells were taken from each trough plate after 2, 4, and 6 h of irradiation, spread on tray plates containing LNG BTB indicator medium, and incubated at 22 °C for 48 h.
The tray plates from several passive and active stackers were moved to the liquid handler in a scheduled fashion where they were spotted with medium and then with irradiated culture from the deep-well trough plates on the deck.
An automated protocol on a robotic workcell (Hughes et al. 2005) was used to spread the 10-μL aliquots from these samples onto the 128 × 96 mm Omni Tray plates.
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