User:Karmella Haynes/Notebook/PcTF Genomics/2012/12/03: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2012/12/03 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
 
Line 8: Line 8:




==mm/dd/yy==
==12/03/12==
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* Line item 1
* Cell culture tasks: stock cells and Carly's transfection


----
----
'''Line item 1'''<br>
'''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 14:28, 8 December 2012

Pc-TF Genomics <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>


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



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/PcTF_Genomics/2012/12/03&oldid=662939"