User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/23: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 27: Line 27:
# well 7 - KAH170/015 (2014)
# well 7 - KAH170/015 (2014)


Abbreviated protocol (from Seron Eaton)
 
Day 1: Abbreviated protocol (from Laura Gonzalez)
* Retrieve frozen aliquots of harvested virus from -80C freezer (just outside TC room)  
* Retrieve frozen aliquots of harvested virus from -80C freezer (just outside TC room)  
* Thaw and quick-spin to get virus off of the inside of lid
* Thaw and quick-spin to get virus off of the inside of lid
* Biohazard set-up: 10 mL Wescodyne waste conical (50 mL), 10 mL Wescodyne rinse
* Biohazard set-up: 10 mL Wescodyne waste conical (50 mL), 10 mL Wescodyne rinse
* Remove 1mL of media from each well (except for K562)
* Add 5ul of 2.2mg/ml Polybrene (thawed) to each well. Note: final concentration of Polybrene in well will be 8μg/ml after 400 μL viral aliquot is added
* Spin at 2250 rpm for 25 minutes
* Place the plates in the incubator (37°C) overnight.




Revision as of 06:45, 28 September 2014

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>


09/23/14

  • Transductions: U2OS, SK-N-SH, K562

Transductions
Note: cells are in standard complete medium

  1. Plate 1: U2OS
  2. Plate 2: SK-N-SH
  3. Plate 4: K562 b (1 mL) - no one was around to consult on whether I should pellet the 2mL and replate

Wells (for all plates) - added 400 μL virus

  1. well 1 - GFP
  2. well 2 - KAH160/015 (2010)
  3. well 3 - KAH160/015 (2014)
  4. well 4 - KAH165/015 (2010)
  5. well 6 - KAH165/015 (2014)
  6. well 7 - KAH170/015 (2014)


Day 1: Abbreviated protocol (from Laura Gonzalez)

  • Retrieve frozen aliquots of harvested virus from -80C freezer (just outside TC room)
  • Thaw and quick-spin to get virus off of the inside of lid
  • Biohazard set-up: 10 mL Wescodyne waste conical (50 mL), 10 mL Wescodyne rinse
  • Remove 1mL of media from each well (except for K562)
  • Add 5ul of 2.2mg/ml Polybrene (thawed) to each well. Note: final concentration of Polybrene in well will be 8μg/ml after 400 μL viral aliquot is added
  • Spin at 2250 rpm for 25 minutes
  • Place the plates in the incubator (37°C) overnight.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/PcTF_Genomics/2014/09/23&oldid=824065"