User:David Benjamin Nyer/Notebook/PcTF breast cancer/2015/11/25

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Summary

BT-474 transfection results and next steps.

BT-474 transfection results

Transfection efficiency very poor throughout, based on fluorescence microscopy. Not going to worry about harvesting and measuring efficiency via flow cytometry.

Next steps: set up two 6-well plates per cell type with 3 cultures per plate. Try these transfection optimization steps:

  1. Transfection followed by 30 minutes centrifugation at 100rpm;
  2. Trypsinization and resuspension in fresh media, followed by transfection so that cells are non-adherent and floating around in lipo-DNA saturated media.



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