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The supernatant was then removed and sample buffer added to the beads.
Wash buffer was removed and sample buffer was added, and then boiled for 5 min at 95°C for immunoblotting.
Media were then removed and sample buffer (80 μL) added to the wells; plates were stored at −70°C.
Sample d-SS1 was treated with 5 mM sodium ascorbate, 1 μM CuCl, and 5 mM NEM at 25 °C for 60 min. Treatment was removed, and sample d-SS1 was washed with the reaction buffer using the cutoff filter.
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Plant residue and gravel were removed and samples were mixed uniformly to form one composite sample.
After 24 h, frames were removed and samples were placed in mixture of water and some lime for 27 days.
After the incubation period the medium was removed and samples were washed two times with 1 mL PBS buffer.
After 48 h, the supernatant liquid of undisturbed samples is carefully removed and samples are thus concentrated to 30 mL for further processing.
At the end of the incubation period (18 h), TSB was removed and samples were stored without culture media at +4°C for 1 or 2 weeks.
After RNAlater infused the samples, it was removed and samples were maintained at −80°C until processing.
All samples were stored at 4°C and the following day RNAlater® was removed and samples transferred to -80°C until further qPCR processing.
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