Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/06/01
|K562 Neon Transfection Results, Day 5||Main project page|
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After transfection on Wednesday the 27th, the K562 cells were expanded on Friday the 29th from 1mL cultures in 12-well plates to 4 mL cultures in 6-well plates. Today I collected more images of the cells using fluorescence microscopy and measured the percentage of fluorescent cells in culture using flow cytometry.
Observations: Lots of cell debris/dead cells in wells 3, 5, and 6. Looks as though method #3 (wells 3 & 6) is particularly hard on the cells. Dead cells have smaller diameter, more complex internal structures compared to live cells.
Transfection efficiency appears low (<5%) for all of the KAH126 samples. Will consult with Dr. Haynes and see about repeating the experiment using higher cell density and higher concentration of plasmid.