Snapfect™ CRISPR Transfection Protocol
SnapfectTM CRISPR Transfection Protocol
Reagent A – Green cap Reagent B – Blue cap
After seeding cells of interest with a 70-80% density overnight in 6 well culture plates under standard growth conditions, the cells are ready to be transfected. The following protocol is for cell transfection for a single well.
- Prepare the transfection agent by combining 10 μL Reagent B and approximately 3 μg of CRISPR/tRNA plasmid in a DNAase/RNAase free microtube and gently agitate. Then allow the solution to incubate at room temp for 30 minutes.
- Refresh the target cell media with standard growth media, then add approximately 5% v/v Reagent A and gently swirl the plate to mix and incubate the cells for 1-5 min under standard conditions.
- Aspirate the growth media containing Reagent A and wash 1X with PBS or other suitable media once the Reagent B transfection agent is prepared.
- After 30 min has lapsed dilute Reagent B by adding 800 μL of SF media.
- Add 810 μL of Reagent B transfection agent to the well and incubate 37°C + 5% CO2 for 10 minutes
- Add 2mL of growth media and further incubate for 24hr to 48hr for results.