Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2014/12/29: Difference between revisions

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(Autocreate 2014/12/29 Entry for Haynes_Lab:Notebook/RNA-seq_of_PcTF_Transfected_U2OS_&_SK-N-SH_cell_lines)
 
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==Goals==
==Goals==
* Place main goals for the day here<br><br>
* Prepare more U2OS medium (McCoy's 5A w/ 10% FBS and P/S antibiotics)
* Passage SKNSH cell line
* Split SKNSH cell line into 6-well plate in preparation for transfection this week
* Passage U2OS cell line
* Split U2OS cell line into 6-well plate in preparation for transfection
** Use very dilute cell concentration to prevent overgrowth of plate
* Split U2OS cell line into 24-well plate to establish growth curve
** Pull from same dilution that's used to seed 6-well plate
** Be very precise when measuring out cell amounts to ensure good growth curve
* Measure initial U2OS cell concentration using flow cytometry
* thaw frozen U2OS stock w/ doxycycline-inducible PcTF gene, begin PS-α<br><br>


==Protocol==
==Protocol==

Revision as of 09:48, 29 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
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Summary

Explain what you're doing today and why. Paragraph format, simple description so somebody reading the notebook has some orientation.

Goals

  • Prepare more U2OS medium (McCoy's 5A w/ 10% FBS and P/S antibiotics)
  • Passage SKNSH cell line
  • Split SKNSH cell line into 6-well plate in preparation for transfection this week
  • Passage U2OS cell line
  • Split U2OS cell line into 6-well plate in preparation for transfection
    • Use very dilute cell concentration to prevent overgrowth of plate
  • Split U2OS cell line into 24-well plate to establish growth curve
    • Pull from same dilution that's used to seed 6-well plate
    • Be very precise when measuring out cell amounts to ensure good growth curve
  • Measure initial U2OS cell concentration using flow cytometry
  • thaw frozen U2OS stock w/ doxycycline-inducible PcTF gene, begin PS-α

Protocol

example protocol

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

Any significant data goes here. Thoughts on what happened, significance of findings, what to do next etc.
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