The aqueous phase was mixed with 100% ethanol and 0.1 volumes of 7.5 M NH4Acetate, incubated at -20°C from 2 hours to overnight (ON), followed by washing with 70% ethanol.
Shortly, a smear of cells from overnight growth on GCB plates was resuspended in 25 µl of lysis solution (50 mM Tris/HCl pH 8.0, 20 mM EDTA, 50 mM NaCl).
Inoculum suspensions were prepared by selecting colonies from overnight growth on nutrient agar plates.
Colonies were picked from overnight growth on the zeocin plate for further PCR confirmation analysis and commercial DNA sequencing analysis.
Bacterial saline suspensions were prepared from overnight cultures on blood agar and adjusted to a 0.5 McFarland turbidity standard.
The isolates were identified via a commercial biochemical identification system using colonies obtained from overnight growth on solid media (API 20NE) (bioMérieux, Marcy l'Etoile, France).
Inocula were prepared from overnight growth on chocolate agar in 5% CO2 at 37°C for NG, from blood agar incubated anaerobically overnight at 37°C for FN, and from blood agar incubated in 5% CO2 at 37°C for 48 h for LB (media from Oxoid, Basingstoke, UK).
A railway mail worker in 1963, Mr. Allison said he came home from New York overnight on the morning of the march, took a nap and washed up.
He also rescues an unconscious young woman (Hannah Gross) from freezing overnight on a park bench, and when she returns the next day with a pie, it's clear something's in store for Noel.
After being rinsed three times during 10 min in 1 ml PT, ovaries were incubated in 500 μl PT + 0.5 μl TOTO-3 (1 mM, Invitrogen, Life Technologies GmbH, Germany) + 0.5 μl SYTOX Orange (5 mM, Invitrogen, Life Technologies GmbH, Germany) and protected from light, overnight (ON) at 4°C.
