293T cells were maintained in DMEM (Dulbecco’s Modified Eagle’s Medium) containing 10% FBS (Fetal Bovine Serum) and 1% PSQ (Penicillin-Streptomycin-Glutamine). LR73 cells were maintained in Alpha-MEM (Alpha’s Modified Eagle’s Medium) containing 10% FBS and 1% PSQ. 293T cells were transfected by using calcium phosphate transfection system (Promega, Madison, WI, USA) and LR73 cells were transfected by using Lipofectamin 2000 (Invitrogen, Waltham, MA, USA) according to the manufacturer’s protocol.
Calcium phosphate transfection system
Manufactured by Promega
Sourced in United States
The Calcium phosphate transfection system is a laboratory equipment used to introduce genetic material, such as plasmid DNA, into cells. It utilizes the formation of calcium phosphate-DNA co-precipitates to facilitate the uptake of the genetic material by the target cells.
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Lab products found in correlation
2 protocols using calcium phosphate transfection system
1
Cell Culture and Transfection Protocols
2
VSV-Pseudotyped HIV-1 Virus Production
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High-titer VSV-pseudotyped recombinant virus stocks were produced in 293T cells by cotransfection of pNL4-3.Luc.R-E- together with the pcDNA3-VSV plasmid encoding the vesicular stomatitis virus G-protein using the calcium phosphate transfection system (Promega, Madison, WI, USA). Supernatants containing virus stocks were harvested 48 h post-transfection, centrifuged to remove cell debris, and stored at -80°C until use. Cell-free viral stock was tested using an enzyme-linked immunoassay for antigen HIV-p24 detection (Innotest HIV-1 antigen mAb; Innogenetic, Ghent, Belgium).
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