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Protocol#
- Aspirate media and wash with PBS
- Incubate with PBS+Tryple for 1min30sec at RT
- Aspirate and wash with PBS
- Add RPMI(+)+Polybrene
- Add particles dropwise (aim to 20x final concentration
Expectations#
- Takes a long time in CMs to get expression compared to HEK cells, around 4 days.
- Non CMs have higher expression of the exogenous protein.
Material#
- PBS+Tryple mix 1to1
- RPMI(+) with Polybrene (7.5uL polybrene/10mL of media); 1.2mL per 12well well (add cat num)
- Viral particles (ideally 30-45x concentrated)
Sanity check#
- Test particles at 1x concentrated on Cal51 or HEK cells
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