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> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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==Goals== | ==Goals== | ||
* | * Determine transfection efficiency of U2OS cells transfected the previous day<br><br> | ||
==Protocol== | ==Protocol== | ||
None<br><br> | |||
==Materials== | ==Materials== | ||
None<br><br> | |||
==Notes== | ==Notes== | ||
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 Previous entry Next entry |
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 NotesComposite image of phase contrast + mCherry fluorescence, adherent U2OS cells: 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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