Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/02/27: 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"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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==Goals==
==Goals==
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* List goals here, in list format
* &#x2713; Measure cell density of K562 culture via flow cytometry
* &#x2713; If cell density is between 500,000 and 1,000,000 cells/mL, transfer to fresh antibiotic-free medium and transfect with lipofectamine, KAH126/MV2 plasmid


<br><br>
<br><br>


==Protocol==
==Protocol==
[http://openwetware.org/wiki/Haynes:ThawingCells example protocol]
[http://openwetware.org/wiki/Haynes:FCMamalian_FluorGene flow cytometry]
 
[http://openwetware.org/wiki/Haynes:TransfectionPlasmid_Lipo transfection with lipofectamine]




<br><br>
<br><br>
==Notes==
==Notes==
Observations, measurements, placeholder info etc. go here.
Well 1: KAH126/MV2, ben's plasmid prep (original concentration 377 ng/μL)<br>
Well 2: KAH126/MV2, Mara's plasmid prep (original concentration 279 ng/μL)<br>
Well 3: negative control, no plasmid


<br><br>
<br><br>


==Results/Conclusions==
==Results/Conclusions==
Data collected from your experiments, significance of findings, future steps etc. all go here. Like the introduction of the page, this should also be in paragraph format and summarize your findings so a new reader can follow along.
 
Culture density is 668,000 cells/mL as measured via flow cytometry.
 
Cells have been transfected and placed in the incubator. Tomorrow: fluorescence microscopy to check transfection efficiency.




Latest revision as of 00:47, 27 September 2017

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

In paragraph format, state what you are going to do today so a reader unfamiliar with your project could understand what is going on.



Goals

  • ✓ Measure cell density of K562 culture via flow cytometry
  • ✓ If cell density is between 500,000 and 1,000,000 cells/mL, transfer to fresh antibiotic-free medium and transfect with lipofectamine, KAH126/MV2 plasmid



Protocol

flow cytometry

transfection with lipofectamine




Notes

Well 1: KAH126/MV2, ben's plasmid prep (original concentration 377 ng/μL)
Well 2: KAH126/MV2, Mara's plasmid prep (original concentration 279 ng/μL)
Well 3: negative control, no plasmid



Results/Conclusions

Culture density is 668,000 cells/mL as measured via flow cytometry.

Cells have been transfected and placed in the incubator. Tomorrow: fluorescence microscopy to check transfection efficiency.



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