User:Karmella Haynes/Notebook/PcTF Genomics/2014/11/15

From OpenWetWare
Jump to navigationJump to search
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>


11/15/14

  • Transfection: KAH126/MV2 into U2OS


Transfection: KAH126/MV2 into U2OS
Strategy: instead of trying to increase transfection efficiency, do a "quasi-subcloning" where transfected cells are separated into smaller wells, and wells showing highest RFP will be selected, pooled, and re-plated.

  • 6-well plate seeded on 11/14/14 (~2.5E5 cells/well)
  • Change medium to p/s-free before transfection (4 mL per well)
  • Use standard Lipofectamine LTX procedure w/ 2000 ng plasmid
  1. KAH126/MV2
  2. KAH126/MV2
  3. no transfection
  • Grow overnight, look for RFP next day
  • Next day: harvest transfected cells and use 1/2 (5.0E5) for 2 different plates
    • 24-well plate - ~2.1E5 cells/well
    • 96-well plate -


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/PcTF_Genomics/2014/11/15&oldid=843958"