5Combine transfection reaction with growth media02:30
6Add transfection mixture to cells02:50
7Incubate cells and observe gene expression03:03
Cell BiologyYouTube (Curated Tutorials)
Transfection Tutorial
Protocol
Difficulty
intermediate
Steps
1
Gather all required reagents and materials
Collect a 12-well plate with Chinese hamster ovary cells, pS40 Campari plasmid DNA (1 microgram per microliter), Fugene transfection reagent, transfection media, and growth media.
▶ 00:44
2
Mix DNA with Fugene reagent
Add DNA to transfection media, then add Fugene reagent, vortex briefly, and incubate the mixture for 5 minutes at room temperature.
▶ 01:25
3
Prepare cells by removing culture media
While the transfection mixture incubates, aspirate the growth media from all wells of the 12-well plate and properly dispose of the aspirator tip.
▶ 01:50
4
Set up untransfected control well
Add 1 milliliter of fresh growth media to one well that will serve as an untransfected control for comparison.
▶ 02:15
5
Combine transfection reaction with growth media
Transfer the entire transfection reaction into fresh growth media using a P1000 pipette, then gently invert the tube about 5 times to mix thoroughly.
▶ 02:30
6
Add transfection mixture to cells
Add 1 milliliter of the mixed transfection reaction to each well intended for transfection, avoiding the untransfected control well.
▶ 02:50
7
Incubate cells and observe gene expression
Return the 12-well plate to a 37°C incubator for 48 hours until green fluorescence from Campari expression becomes visible in the transfected cells.
▶ 03:03
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