Home Protocols Combination of TransPass D1 or TransPass D2 & TransPass V (M2561)

Combination of TransPass D1 or TransPass D2 & TransPass V (M2561)

Protocol

  1. In a tube, mix plasmid(s) and/or siRNA in serum free medium.
  2. Add TransPass D1 or TransPass D2 to the above mixture.
  3. Incubate at room temperature for 25-30 minutes.
  4. Add TransPass V to the incubated mixture.
  5. (Optional) Incubate at room temperature for 15 minutes.
  6. Add the transfection complex mixture to cells (in complete growth media).
  7. Incubate at 37°C, 5% CO2 for 24 hours before replacing media. 
    Note: Replace media every day, if longer incubation period is required. 

    Table 1:     Transfection complex mixtures using TransPass D1 or TransPass D2 & TransPass V
    Culture Vessel Surface
    Area    		 Volume
    of Plating
    Medium
    (per well)    		 *Total DNA in
    Serum-free
    Medium Volume
    (per well)    		 TransPass
    D1 or D2
    (per well)    		 TransPass V
    (per well)
    96 well 0.32 cm2 100 µ 0.1 µg in 10 µl 0.1–0.3 µl 0.1–0.9 µl
    48 well 0.95 cm2 250 µ 0.3 µg in 25 µl 0.3–0.9 µl 0.3–2.7 µl
    24 well 1.9 cm2 500 µl 0.7 µg in 50 µl 0.7–2.1 µl 0.7–6.3 µl
    12 well 3.8 cm2 1 ml 1.5 µg in 150 µl 1.5-4.5 µl 1.5–13.5 µl
    35 mM or 6 well 9.5 cm2 2 ml 3 µg in 250 µl 3–9 µl 3–27 µl
    60 mM dish 21 cm2 5 ml 6 µg in 500 ml 6–18 µl 6–54 µl
    100 mM dish 55 cm2 15 ml 18 µg in 1 ml 18–54 µl 18–162 µl<
    *For plasmid and siRNA transfection, we recommend 20–100 nM siRNA per well in addition to the suggested DNA amount and the 1:1, 1:2 or 1:3 ratio (TransPass transfection reagent: TransPass V).