ESO wikiadmin: Created page with "{| class="wikitable" |+ !Materials ! |- | |DMEM + 10% CS |- | |Puromycin (stock solution 5 mg/ml) |} === Procedure === # 1x10⁴ 293T cells are seeded in a 24-well plate ~12 h before transduction (5 wells) # 1µl, 2µl or 5µl, respectively, of the virus-concentrate are added on top of the cells, 2 wells remain untreated # incubation over night at 37°C and 5% CO₂ # 24 h after transduction the medium is replaced by 1 ml fresh medium containing 10% C..."

2025-02-15T17:48:22Z

Created page with "{| class="wikitable" |+ !Materials ! |- | |DMEM + 10% CS |- | |Puromycin (stock solution 5 mg/ml) |} === Procedure === # 1x104 293T cells are seeded in a 24-well plate ~12 h before transduction (5 wells) # 1µl, 2µl or 5µl, respectively, of the virus-concentrate are added on top of the cells, 2 wells remain untreated # incubation over night at 37°C and 5% CO2 # 24 h after transduction the medium is replaced by 1 ml fresh medium containing 10% C..."

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{| class="wikitable"
|+
!Materials
!
|-
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|DMEM + 10% CS
|-
|
|Puromycin (stock solution 5 mg/ml)
|}

=== Procedure ===

# 1x104 293T cells are seeded in a 24-well plate ~12 h before transduction (5 wells)
# 1µl, 2µl or 5µl, respectively, of the virus-concentrate are added on top of the cells, 2 wells remain untreated
# incubation over night at 37°C and 5% CO2
# 24 h after transduction the medium is replaced by 1 ml fresh medium containing 10% CS and 0,4 µg/ml puromycin (one of the wells harbouring untransduced cells will not be treated with puromycin)
# after 4-5 days the cell number is defined and the viral titer (viral particles / µl) is calculated by counting still viable cells

Transduction and titration of lentiviral supernatant - Revision history