User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/10

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09/10/14

  • Lenti Transfection: Day 2 - change out medium (LaBaer lab)


Lenti Transfection: Day 2 - change out medium
Note: At this point, cells are producing virus. Follow extra biosafety precautions.

  • Medium change 1 - 10:00 am
  1. PPE note: Wear 2x pairs of gloves. Dispose of and replace outer gloves as needed.
  2. In the BS cabinet, set up 2x 50 mL conicals of 10 mL Wescodyne (brown) germicidal reagent. Use one for discarding viral medium, one for rinsing pipettes.
  3. Checked GFP control (sample 7) under scope. Nearly 100% expression. (Note: scope had no RFP filter set)
  4. For each well (one at a time)...
    1. Removed 1.5 mL growth medium, dispensed into Wescodyne, rinsed pipette in Wesc., discarded pipette
    2. Slowly and carefully added 2 mL fresh growth medium, rinsed pipette in Wesc., discarded pipette



Reagent Volume
reagent 1 # μL
reagent 2 #
reagent 3 #
reagent 4 #
dH2O #
  # μL

--> Reaction conditions



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