Team:Tsinghua/Notebook/Protocol/293T transfection

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Notebook: Protocol

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Transient 293T Transfection with MegaTran 1.0 (OriGene)

1.Preparation of 293T cells: Plate ~2x105 adherent cells per well 24h prior to transfection.

2.Preparation of the MegaTran 1.0/DNA Complex:

Dilute 1ug of DNA in 100ul of Opti-MEM I. Vortex gently.
Add 3ul of MegaTran 1.0 to the diluted DNA (not the reverse order!) and vortex the solution immediately for 10 seconds.
Incubate for 10 minutes at room temperature.

3.Transfection

Add the Megatran 1.0/DNA mixture from step 2 to each well (already containing about 2ml culture medium) gently.
Gently rock the plate to achieve even distribution of the complexes.
Incubate at 37oC for 24-48h.

 

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