Pseudovirus Concentration Protocol

Pseudoviruses were concentrated as described before (Mykytyn et al., 2021 (link)) on a 10% sucrose cushion (10% sucrose, 15 mM Tris–HCl, 100 mM NaCl, 0.5 mM EDTA) for 1.5 hr at 20,000 x g at 4°C. Supernatant was decanted and pseudoviruses resuspended in Opti-MEM I (1X) + GlutaMAX (Gibco) to achieve 100-fold concentration.

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Lamers M.M., Mykytyn A.Z., Breugem T.I., Wang Y., Wu D.C., Riesebosch S., van den Doel P.B., Schipper D., Bestebroer T., Wu N.C, & Haagmans B.L. (2021). Human airway cells prevent SARS-CoV-2 multibasic cleavage site cell culture adaptation. eLife, 10, e66815.

Publication 2021

Opti-MEM I (1X) GlutaMAX

Edta Nacl Sucrose Tris

Corresponding Organization :

Other organizations : University of Illinois Urbana-Champaign, Invitae (United States)

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Variable analysis

independent variables

Centrifugation at 20,000 x g for 1.5 hr at 4°C

dependent variables

Concentration of pseudoviruses

control variables

10% sucrose cushion (10% sucrose, 15 mM Tris–HCl, 100 mM NaCl, 0.5 mM EDTA)
Resuspension of pseudoviruses in Opti-MEM I (1X) + GlutaMAX (Gibco)

controls

No positive or negative controls were explicitly mentioned.

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