Haynes Lab:Notebook/Synthetic Biology and Bioinformatics for Predictable Control of Therapeutic Gene2/2012/12/18: Difference between revisions

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(Autocreate 2012/12/18 Entry for Haynes_Lab:Notebook/Synthetic_Biology_and_Bioinformatics_for_Predictable_Control_of_Therapeutic_Gene2)
 
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==Entry title==
==Cells in Trizole==
* Insert content here...
* Cells in trizole at 80°C
'''K562'''
#Expand cells to 10cm petri dishes for 10mL of cells
#For the 6 plates, transfer 1mL cells to 2mL tubes 4 times (8mL total)
#Spin tubes at room temperature @ 3000 RPM for 5 min
#Dispose of supernatant
#Add 500μL of trizol to each pellet
#Store @ -80°C
'''SK-N-SH'''
#From the 6 well plate aspirate off the growth media
#Add 500μL of trizol directly to the 6 well plate
#Let sit for 5 min @ room temperature under the hood
#Use 100m0mL pipet to scrape and pipet up and down to break up cells
#Transfer 500μL into each tube




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Cells in Trizole

  • Cells in trizole at 80°C

K562

  1. Expand cells to 10cm petri dishes for 10mL of cells
  2. For the 6 plates, transfer 1mL cells to 2mL tubes 4 times (8mL total)
  3. Spin tubes at room temperature @ 3000 RPM for 5 min
  4. Dispose of supernatant
  5. Add 500μL of trizol to each pellet
  6. Store @ -80°C

SK-N-SH

  1. From the 6 well plate aspirate off the growth media
  2. Add 500μL of trizol directly to the 6 well plate
  3. Let sit for 5 min @ room temperature under the hood
  4. Use 100m0mL pipet to scrape and pipet up and down to break up cells
  5. Transfer 500μL into each tube


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