Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/01/05
|RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines||Main project page|
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Today I'll be checking up on the transfected U2OS cells, hopefully harvesting them for RNA sequencing and quantification. I'll also be checking on the SKNSH culture in the 6-well plate and advancing the KAH126/U2OS cell line.
Observations, measurements, placeholder info etc. go here
Phase / mCherry composite images of transfected U2OS cultures below.
Regardless, I'm not optimistic about getting high quality RNA data from these samples, as cells that are not adherent at this stage of the culture are not likely to be viable or biologically active. Future work might entail splitting the samples 1 day after transfection, in case overcrowding is the issue.
After speaking with Dr. Haynes we've decided to halt work on the transiently transfected U2OS cells and will not collect RNA from the cells or measure their fluorescence via flow cytometry. I'll continue to advance the KAH126/U2OS cell line, passage the unmodified U2OS line and look into using nucleofection on the U2OS cell line from another bioengineering lab.