Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/05/06

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Overview

A U2OS culture in a 6-well plate was transfected with the following plasmids:

  • Wells 1 & 2: pMaxGFP
  • Wells 3 & 4: KAH187-MV8 (CFP with a CMV promoter)
  • Wells 5 & 6: KAH126-MV2 (PcTF-RFP with a CMV-tet promoter (should be always on))

Today I took images of the six wells. The CFP lamp does not appear to be functional on the microscope, but there is sufficient overlap of fluorescence between CFP and GFP that I could image using GFP for the samples treated with KAH187-MV8.

Transfection Results

pMaxGFP:

2015-05-06 Well 1 pMaxGFP phase GFP 10x.jpg 2015-05-06 Well 2 pMaxGFP phase GFP 10x.jpg


KAH187-MV8:

2015-05-06 Well 3 KAH187-MV8 phase GFP 10x.jpg 2015-05-06 Well 4 KAH187-MV8 phase GFP 10x.jpg


KAH126-MV2:

2015-05-06 Well 5 KAH126-MV2 phase mCherry 10x.jpg 2015-05-06 Well 6 KAH126-MV2 phase mCherry 10x.jpg

Positive controls worked fine, but still no transfection with KAH126 plasmid. The mystery continues...