general summary

100 uL Mirus reagent
2 ug DNA(up to 20 uL DNA solution)
1 – 5 million cells/mL Lonza Knowledge Center

Protocol

• Warm medium in flask and coverslip-bottom dish
• Mix DNA and Mirus reagent; warm up
• Trypsinize cells:
  • remove medium
  • rinse with warm PBS
  • add 0.5 mL trypsin
  • incubate till all cells are in suspension (at least 3 min)
  • add 1 mL COLD medium
  • mix well by pipetting up and down
• transfer all to sterile 2 mL tube
• spin 500 x g 3 minutes
• resuspend cells in Mirus/DNA solution
• put all into sterile cuvette, close
• Put cuvette into Nucleofector
• Choose program¹
• Press the button
• Use special dropper to transfer one drop to the coverslip dish and the rest to the waiting flask.

Mirus nucleofection kit