Creativity

Innovation

Originality

Imagination

 

Salient

Salient is an excellent design with a fresh approach for the ever-changing Web. Integrated with Gantry 5, it is infinitely customizable, incredibly powerful, and remarkably simple.

Download

Transfection of mammalian tissue culture cells

 

FuGENE 6 (Roche)

General considerations:

  • Warm to room temperature before use
  • Mix before use
  • Do not aliquot
  • Always pipet into serum-free medium, not into empty tubes
  • Initial testing
  • In certain cells antibiotics or serum present in the medium may have adverse effects
  • Use FuGENE (µl):DNA (µg) amounts of 3:2, 3:1 and 6:1

Preparation of complex

  1. Pipette serum free medium into a tube, followed by the FuGENE6. Tap to mix.
  2. Add the DNA.
  3. Tap the tube to mix. DO NOT VORTEX.

Transfection procedure

  1. Dropwise, add the complex to the cells, distributing around the well. Swirl to ensure even dispersal.
  2. Return the cells to the incubator.

Table 1. Recommended amounts of components.

Type of dish or plate Surface area (cm2) Total media volume (ml) Dilution volume (µl) Starting volume of FuGENE 6 (µl) Starting mass of DNA (µg)

60 mm
35 mm
6-well
12-well
24-well
96-well

21
8.0
9.4
3.8
1.9
0.3

4
2
2
1
0.5
0.1
100
100
100
50
20
5
6.0
3.0
3.0
1.5
0.6
0.15

2.0
1.0
1.0
0.5
0.2
0.05