Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/06/01
K562 Neon Transfection Results, Day 5 | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
OverviewAfter 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. Fluorescence MicroscopyObservations: 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. Flow Cytometry
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.
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