Difference between revisions of "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)
 
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==mm/dd/yy==
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==12/03/12==
 
<!-- Precede finished items with a checkmark &#x2713; -->
 
<!-- Precede finished items with a checkmark &#x2713; -->
* Line item 1
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* Cell culture tasks: stock cells and Carly's transfection
  
 
----
 
----
'''Line item 1'''<br>
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'''U2OS'''<br>
> Samples
+
* 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
  
{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
 
|-valign="top"
 
| <u>Reagent</u> || <u>Volume</u>
 
|-
 
| reagent 1 || # μL
 
|-
 
| reagent 2 || #
 
|-
 
| reagent 3 || #
 
|-
 
| reagent 4 || #
 
|-
 
| dH<sub>2</sub>O || #
 
|-
 
| &nbsp; || # μL
 
|}
 
  
--> Reaction conditions
+
'''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
  
  

Revision as of 13:28, 8 December 2012

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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