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Plasmid Transfection in 293T Cells: Principles and Hands-on Protocol

🚨 Failure Case Library (4) + Submit your own case

severe
Bright fluorescence but Western blot shows very low expression
GFP fluorescence is obvious under microscope, but the corresponding band on Western blot is faint or near-invisible.
💡 5 · ✓ 5
moderate
mRNA Transfection Toxicity
Cell toxicity observed during mRNA transfection experiments with MessengerMAX reagent
💡 5 · ✓ 7
moderate
Co-transfection ratio imbalance — uneven dual-plasmid expression
Of two co-transfected plasmids, only one expresses strongly; the fraction of double-positive cells is low.
💡 5 · ✓ 4
moderate
Abnormal fluorescence localization (aggregates / wrong compartment)
Fluorescence forms aggregates inside cells, or localizes to a compartment different from the expected native pattern.
💡 4 · ✓ 4
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