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.
PPE note: Wear 2x pairs of gloves. Dispose of and replace outer gloves as needed.
- Medium change 1 - 10:00 am
- In the BS cabinet, set up 2x 50 mL conicals of 10 mL of 10x Wescodyne (brown) germicidal detergent. Use one for discarding viral medium, one for rinsing pipettes.
- Checked GFP control (sample 7) under scope. Nearly 100% expression. (Note: scope had no RFP filter set)
- For each well (one at a time)...
- Removed 1.5 mL growth medium, dispensed into Wescodyne, rinsed pipette in Wesc., discarded pipette
- Slowly and carefully added 2 mL fresh growth medium, rinsed pipette in Wesc., discarded pipette
- Final volume ~3 mL
- Placed plates back into 37°C incubator.
- Let discarded medium/Wesc. sit for 15 min.
- Wiped down BS cabinet surface with Microbiocide.
- Aspirated off discarded waste, then rinse solution.
- Medium change 2 - 4:00 pm
- Same biosafety/ discard set-up as before
- For each well (one at a time)...
- Removed 2.5 mL growth medium, dispensed into Wescodyne, rinsed pipette in Wesc., discarded pipette
- Slowly and carefully added 5.5 mL fresh growth medium, rinsed pipette in Wesc., discarded pipette
- Placed plates back into 37°C incubator.
- Same biosafety/ discard procedure as before
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