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

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines</span>
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==Goals==
==Goals==
* Place main goals for the day here<br><br>
* Determine transfection efficiency of U2OS cells transfected the previous day<br><br>


==Protocol==
==Protocol==
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==Materials==
==Materials==
* List materials here, to be gathered before the experiment
None<br><br>
* Consumables such as media, antibiotics, DNA, kits etc.<br><br>


==Notes==
==Notes==
Observations, measurements, placeholder info etc. go here<br><br>
Composite image of phase contrast + mCherry fluorescence, adherent U2OS cells:<br><br>
 
[[Image:2014-12-17_U2OS_cells_transfected_with_mCherry-PcTF.jpg|900px|]]


==Results/Conclusions==
==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.<br><br>
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.<br><br>
*'''[[User:David Benjamin Nyer|David Benjamin Nyer]] 18:38, 17 December 2014 (EST)''':
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__NOTOC__
__NOTOC__

Latest revision as of 00:36, 27 September 2017

RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines Main project page
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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

  • Determine transfection efficiency of U2OS cells transfected the previous day

Protocol

None

Materials

None

Notes

Composite image of phase contrast + mCherry fluorescence, adherent U2OS cells:

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.
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