general summary
100 uL Mirus reagent
2 ug DNA(up to 20 uL DNA solution)
1 - 5 million cells/mL
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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.
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Cell type Program # B16 P-031 3t3 U-030 HCT116 D-032 HeLa I-013 LLCPk-1 X-001 MEF T-020, A-023
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