Viral transduction of mammalian cells via spinfection: Difference between revisions
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Created page with "{| class="wikitable" |+ !Materials ! |- | |Virus containing supernatant |- | |1000x Polybrene (Hexadimethrine bromide) in PBS (8mg/ml) 1*106 suspension cells or 1*105 adherent cells |- | |24 well plate (for suspension cells) 6 well plate (for adherent cells) |} === Procedure === ---- # 1 ml medium containing 1*105 or1*106 cells are added into 6 or 24 well plate. # Add 2 µl 1000x Polybrene # Add 1ml of virus supernatant or a corresponding dilution of concentrated virus..." |
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# 1 ml medium containing 1* | # 1 ml medium containing 1*10<sup>5</sup> or1*10<sup>6</sup> cells are added into 6 or 24 well plate. | ||
# Add 2 µl 1000x Polybrene | # Add 2 µl 1000x Polybrene. | ||
# Add 1ml of virus supernatant or a corresponding dilution of concentrated virus to the well | # Add 1ml of virus supernatant or a corresponding dilution of concentrated virus to the well. | ||
# Perform spinfection at 800 g for 1 h at room temperature, disable the brakes of the centrifuge and spin with acceleration = 1 and deceleration = 1. | # Perform spinfection at 800 g for 1 h at room temperature, disable the brakes of the centrifuge and spin with acceleration = 1 and deceleration = 1. | ||
# Change medium after 1-2 days | # Change medium after 1-2 days. | ||
Latest revision as of 18:50, 15 February 2025
| Materials | |
|---|---|
| Virus containing supernatant | |
| 1000x Polybrene (Hexadimethrine bromide) in PBS (8mg/ml) 1*106 suspension cells or 1*105 adherent cells | |
| 24 well plate (for suspension cells) 6 well plate (for adherent cells) |
Procedure
- 1 ml medium containing 1*105 or1*106 cells are added into 6 or 24 well plate.
- Add 2 µl 1000x Polybrene.
- Add 1ml of virus supernatant or a corresponding dilution of concentrated virus to the well.
- Perform spinfection at 800 g for 1 h at room temperature, disable the brakes of the centrifuge and spin with acceleration = 1 and deceleration = 1.
- Change medium after 1-2 days.