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Lower than expected transfection efficiency
My transfection efficiency has recently dropped even though I am using the same cells and reagents. What are some possible causes?

  • Verify that cells are contaminant-free.  Mycoplasma contamination can cause a suppression of transfection efficiency.
  • “Older” cells transfect with less efficiency.  For best results use cells at lowest passage number possible.
  • If working with self-isolated primary cells, even slight differences in the isolation protocol can cause differences in transfection efficiency.
  • Any differences in cell handling prior to transfection can cause differences in transfection efficiency.
  • Verify cell confluency or density is consistent from experiment to experiment.  Recommend 70-90% confluency or cell density of 0.6-0.8 million suspension cells/ml at the time of transfection.  Variation, higher or lower, can influence observed efficiency.
  • Recommend that the cells are allowed to recover from cryopreservation for at least one passage prior to transfection
Date Created10/15/2013 03:32 PM
Date Updated03/27/2014 08:43 PM

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