Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2014/12/17: Difference between revisions
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==Goals== | ==Goals== | ||
* Determine transfection efficiency | * Determine transfection efficiency of U2OS cells transfected the previous day<br><br> | ||
==Protocol== | ==Protocol== | ||
Revision as of 16:16, 17 December 2014
 RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines
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SummaryToday we're checking the transfected U2OS cells in the 6 well plates for transfection efficiency. This is done by putting the sample under the fluorescence microscope and observing mCherry fluorescence. mCherry is a fluorescent marker tagged to the Pc-TF protein expressed in the plasmid. The more cells with fluorescent red nuclei (Pc-TF has a nuclear colocalization tag), the greater the transfection efficiency. Goals
ProtocolNone MaterialsNone NotesObservations, measurements, placeholder info etc. go here Results/ConclusionsTransfection efficiency appears to be low, <10%. Could be due to low-quality plasmid DNA, or operator error. Going to repeat the transfection procedure, starting by splitting U2OS cells into 6-well plates tomorrow.
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