Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/02/09

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Summary

Following up on the transfection of SKNSH cells last Thursday, today I'm going to image them again and make a decision on whether to harvest for RNA-seq. Also need to start a culture of K-562 cells in preparation for transfecting them with PcTF-RFP.



Goals

  • ✓ Fluorescence microscopy of SKNSH cells transfected with PcTF-RFP
  • ✓ If cells look good, harvest RNA using RNEasy method
  • ✓ Thaw K562 cells, start culture



Protocol

TRIzol/RNeasy protocol

Thawing cells




Notes

Observations, measurements, placeholder info etc. go here.



Results/Conclusions

Fluorescence microscopy images:

Well 1 Well 2

Well 3 (negative control)

Harvested the cells (2 biological replicates, 1 control), RNA is in the -80°C freezer.

K562 culture is in PS-α, in the incubator.

Potential plasmids to use as positive control in transfection/nucleofection: KAH133 (mCherry in MV2), KAH187 (CFP in MV8)