Author: Peña, José; Chen-Harris, Haiyin; Allen, Jonathan E.; Hwang, Mona; Elsheikh, Maher; Mabery, Shalini; Bielefeldt-Ohmann, Helle; Zemla, Adam T.; Bowen, Richard A.; Borucki, Monica K.
Title: Sendai virus intra-host population dynamics and host immunocompetence influence viral virulence during in vivo passage Document date: 2016_4_9
ID: z7f720dj_16
Snippet: For SeV single plaque amplification, LLC-MK2 cell monolayers were seeded in six-well plates and infected with 100 ul of BAL from infected mice and incubated at 37 C for 1 h. Wells were overlaid with 3 ml of 2Â MEM (Life Technologies, Gibco) with 100 IU/ml Penicillin/100 mg/ml Streptomycin (Gibco), 2% BSA (Gibco), 2 mg/ml of TPCK trypsin (Sigma), and 2% agarose. Live plaques were visualized by adding 300 ll MTT solution (1/10 volume of agarose pl.....
Document: For SeV single plaque amplification, LLC-MK2 cell monolayers were seeded in six-well plates and infected with 100 ul of BAL from infected mice and incubated at 37 C for 1 h. Wells were overlaid with 3 ml of 2Â MEM (Life Technologies, Gibco) with 100 IU/ml Penicillin/100 mg/ml Streptomycin (Gibco), 2% BSA (Gibco), 2 mg/ml of TPCK trypsin (Sigma), and 2% agarose. Live plaques were visualized by adding 300 ll MTT solution (1/10 volume of agarose plug: 5 mg/ml tetrazolium sallt, MTT (Sigma) in PBS filtered. Single plaques were collected and resuspended in 1 ml of unsupplemented DMEM (Life Technologies) overnight. Isolated plaques were then amplified in LLC-MK2 cells and supernatant harvested and stored at À80 C on day 7.
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