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Transfection Reagent Selector Kit Handbook

well in 2 ml of appropriate growth medium containing serum and antibiotics. (For suspension cells, seed 1.03.5 x 106 cells in 1.6 ml of medium on the day of transfection.)

  • Incubate cells at 37C and 5% CO2 in an incubator. The cells should be 4080% confluent on the day of transfection.
    • Before transfection we recommend to:
    • Check the cells do they look healthy?
    • Check and note the time between seeding and transfection - this should be kept constant in future experiments.
    • Check and note the confluency of the cells this should be kept constant in future experiments.
    1. Label twenty-two 1.5-ml microfuge tubes 111 for Effectene Reagent and 1222 for SuperFect Reagent.

    2. Label six 1.5-ml microfuge tubes AF for mastermix preparation: AC for DNA-Enhancer mastermix preparation for Effectene Reagent and DF for DNA mastermix preparation for SuperFect Reagent.

      Transfection-complex formation
      The following steps for complex formation refer to the pipetting scheme outlined in Figure 2.

    3. DNA-Enhancer mastermixes for Effectene Reagent
      IMPORTANT: It is important to prepare mastermixes AC in the order specified,
      since direct mixing of the DNA with the Enhancer may lead to precipitation.
      Mastermix A: Dilute 0.8 g of DNA in Buffer EC and add 6.4 l of Enhancer. The
      final volume should be 400 l.
      Mastermix B: Dilute 1.6 g of DNA in Buffer EC and add 12.8 l
      '"/>


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