User:Karmella Haynes/Notebook/PcTF Genomics/2012/12/03

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(Autocreate 2012/12/03 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
Current revision (17:29, 8 December 2012) (view source)
(12/03/12)
 
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==mm/dd/yy==
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==12/03/12==
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* Line item 1
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* Cell culture tasks: stock cells and Carly's transfection
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'''Line item 1'''<br>
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'''U2OS'''<br>
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> Samples
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* Froze down 5 vials of U2OS (10% DMSO in Tet-free  FBS), using 100% confluent 75 cm flask
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** Note: Had Rene transfer from -80°C to -150°C on Thursday
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* Seeded 1:10 and 1:20 75 cm flasks from the small back-up flask
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'''SK-N-SH'''
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* Froze down 4 vials of SK-N-SH (10% DMSO in Tet-free  FBS), using 90% confluent 75 cm flask
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** Note: Had Rene transfer from -80°C to -150°C on Thursday
 +
* Seeded 1:5 75 cm flask from 1/5 of the original 75 cm flask
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* These cells don't seem to be very robust
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'''K562'''
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* Seeded 1:10 75 cm flask (passage 5)
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{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
 
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|-valign="top"
 
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| <u>Reagent</u> || <u>Volume</u>
 
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|-
 
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| reagent 1 || # μL
 
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|-
 
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| reagent 2 || #
 
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|-
 
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| reagent 3 || #
 
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|-
 
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| reagent 4 || #
 
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|-
 
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| dH<sub>2</sub>O || #
 
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|-
 
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| &nbsp; || # μL
 
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|}
 
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--> Reaction conditions
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'''K562, Carly's transfection'''
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* Carly observed RFP signal 24 hours after transfection for all three DNA+ samples; no signal in the three negative controls
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* Pelleted (individually) each of the 6 samples (from 6-well dish) at 1000 rpm/ 3 min.
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* Resuspended in growth medium + pen/strep
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* Plated in a total of 10 mL medium + pen/strep in 10 cm petri dishes

Current revision

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12/03/12

  • Cell culture tasks: stock cells and Carly's transfection



U2OS

  • Froze down 5 vials of U2OS (10% DMSO in Tet-free FBS), using 100% confluent 75 cm flask
    • Note: Had Rene transfer from -80°C to -150°C on Thursday
  • Seeded 1:10 and 1:20 75 cm flasks from the small back-up flask


SK-N-SH

  • Froze down 4 vials of SK-N-SH (10% DMSO in Tet-free FBS), using 90% confluent 75 cm flask
    • Note: Had Rene transfer from -80°C to -150°C on Thursday
  • Seeded 1:5 75 cm flask from 1/5 of the original 75 cm flask
  • These cells don't seem to be very robust


K562

  • Seeded 1:10 75 cm flask (passage 5)


K562, Carly's transfection

  • Carly observed RFP signal 24 hours after transfection for all three DNA+ samples; no signal in the three negative controls
  • Pelleted (individually) each of the 6 samples (from 6-well dish) at 1000 rpm/ 3 min.
  • Resuspended in growth medium + pen/strep
  • Plated in a total of 10 mL medium + pen/strep in 10 cm petri dishes



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