Part of the supernatants was inoculated to shM2-MDCK for next passage of the virus culture, another part was subjected for plaque assay to determine if potential siRNA-resistant virus appeared.
Then, supernatants were collected and used for the next passage of cells.
The lungs were combined and homogenized in phosphate-buffered saline (PBS)–penicillin-streptomycin, the homogenates were centrifuged at 3,000 g for 10 min, and the supernatant was used for the next passage.
Also, in writing for an orchestra, one of Bruckner's favorite devices was to give a passage to one group of instruments and then abruptly switch to a contrasting group for the next passage.
Re-isolations were pooled, frozen, and used to generate the inoculum for the next passage.
After reaching 70 80% confluency, the total number of cells in each culture was determined and a fraction of them was re-plated for a next passage under the same conditions as before.
The supernatant from each passage became the inoculum for the next passage.
Hemagglutination (HA) testing was done with harvested material to define the starting material for the next passage.
Infected blood of either mouse reaching adequate parasitaemia (several percent) was collected 1 week later by cardiac puncture and used for the next passage.
