Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/02/02

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I'm checking on cultures today, nothing special.


  • Monitor cultures
  • If SKNSH samples in 6-well plates show fluorescence, harvest RNA using TRIzol/RNEasy method


example protocol


Observations, measurements, placeholder info etc. go here.


Fluorescence microscopy images of SKNSH samples, 4 days post-transfection:

Well 1 Well 2

Well 3 Well 4

Little to no fluorescence in any of the treated samples. I'm calling this run a dud, will try again with lower concentration of cells. Also need to look into the possibility of supercoiled plasmid resulting in lowered efficiency, and if so best treatment options.

Looked at the other cultures, only U2OS in flask is close to ready. Will passage U2OS tomorrow.