Home Protocols Combination of TransPass R1 & TransPass V (M2561)

Combination of TransPass R1 & TransPass V (M2561)

Protocol

  1. In Tube #1, mix plasmid(s) and/or siRNA together.
  2. In Tube #2, add TransPass R1 to serum-free medium.
  3. Incubate the content of Tube #2 at room temperature for 15 minutes.
  4. Add the content of Tube #1 to Tube #2.
  5. Incubate at room temperature for 15 minutes.
  6. Add TransPass V.
  7. Add the transfection complex mixture to cells (in complete growth media).
  8. Incubate at 37°C, 5% CO2 for 24 hours before replacing media.
    Note: Replace media every day, if longer incubation period is required.

    Table 2: Transfection complex mixtures using TransPass R1 & TransPass V
    Culture Vessel Surface
    Area    		 Volume
    of Plating
    Medium
    (per well)    		 *Total DNA in
    Serum-free
    Medium Volume
    (per well)    		 TransPass R1
    (per well)    		 TransPass V
    (per well)
    96 well 0.32 cm2 100 µ 0.1 µg in 10 µl 0.1–0.3 µl 0.3–1.5 µl
    48 well 0.95 cm2 250 µ 0.3 µg in 25 µl 1–1.5 µl 1–4.5 µl
    24 well 1.9 cm2 500 µl 0.7 µg in 50 µl 2–3 µl 2–9 µl
    12 well 3.8 cm2 1 ml 1.5 µg in 150 µl 4-5 µl 4–15 µl
    35 mM or 6 well 9.5 cm2 2 ml 3 µg in 250 µl 10–12.5 µl 10–37.5 µl
    60 mM dish 21 cm2 5 ml 6 µg in 500 ml 20–28 µl 20-84 µl
    100 mM dish 55 cm2 15 ml 18 µg in 1 ml 52–73 µl 52–219 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).