Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2014/12/17: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 13: Line 13:


==Protocol==
==Protocol==
[http://openwetware.org/wiki/Haynes:ThawingCells example protocol]<br><br>
None<br><br>


==Materials==
==Materials==

Revision as of 16:15, 17 December 2014

RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines <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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Summary

Today 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

  • Place main goals for the day here

Protocol

None

Materials

  • List materials here, to be gathered before the experiment
  • Consumables such as media, antibiotics, DNA, kits etc.

Notes

Observations, measurements, placeholder info etc. go here

Results/Conclusions

Transfection 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.