For production of infectious HIV-1, HEK293T cells were transfected with a plasmid encoding full-length HIV-1NL4.3 by calcium-phosphate DNA precipitation (Takara Bio Europe). Virus-containing culture supernatants were harvested 48–72 h post transfection, filtered using 0.45 μm filters (VWR International), and concentrated via ultracentrifugation through a 20% sucrose cushion (Sigma-Aldrich). The infectious titer of the virus stock was defined by a β-galactosidase-based blue cell assay using TZM-bl cells46 (link).
Siddharta A., Pfaender S., Malassa A., Doerrbecker J., A.n.g.g.a.k.u.s.u.m.a., Engelmann M., Nugraha B., Steinmann J., Todt D., Vondran F.W., Mateu-Gelabert P., Goffinet C, & Steinmann E. (2016). Inactivation of HCV and HIV by microwave: a novel approach for prevention of virus transmission among people who inject drugs. Scientific Reports, 6, 36619.
Other organizations :
Medizinische Hochschule Hannover, Center for Experimental and Clinical Infection Research, Helmholtz Centre for Infection Research, Essen University Hospital, National Development and Research Institutes
Transfection of HEK293T cells with a plasmid encoding full-length HIV-1NL4.3
dependent variables
Infectious titer of the virus stock
control variables
Calcium-phosphate DNA precipitation method for transfection
Virus-containing culture supernatants harvested 48–72 h post transfection
Filtration of virus-containing supernatants using 0.45 μm filters
Concentration of virus via ultracentrifugation through a 20% sucrose cushion
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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