Snapfect™ CRISPR Transfection Protocol

Snapfect^TM 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.

1. 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.
2. 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.
3. Aspirate the growth media containing Reagent A and wash 1X with PBS or other suitable media once the Reagent B transfection agent is prepared.
4. After 30 min has lapsed dilute Reagent B by adding 800 μL of SF media.
5. Add 810 μL of Reagent B transfection agent to the well and incubate 37°C + 5% CO₂ for 10 minutes
6. Add 2mL of growth media and further incubate for 24hr to 48hr for results.

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