Electroporation of A1 cells

Materials:

Ice

4 mM cuvettes, N+1 cuvettes

Steinberg's, at 4 C.

Square pulse electroporator (we use the BTX 830 Squarporator)

1. Put cuvettes on ice, and cool down Steinberg's on ice
2. Trypsinize cells as normal, stop reaction with high serum media and spin down at 1000 rpm, 3 minutes
3. Wash pellet with Steinberg's solution and resuspend in 300 µl of Steinberg's, keep on ice
4. Before electroporation, make sure to electroporate a cuvette with no cells to discharge device
5. Electroporate cells in following conditions and put back on ice: Myoblasts: 100V, 35 msec, 5 pulses, Myotubes: 75 V, 35 msec, 5pulses
6. Plate cells as normal

Steinberg's

1x	FW (MW)	5x/500 ml
58 mM NaCl 58.44 8.47 g 0.67 mM KCl 74.56 1.68 ml (1M) 0.44 mM Ca(NO3) 236.20 1.1 ml (1M) 1.3 mM MgSO4 246.47 3.25 ml (1M) 4.6 mM Tris pH 7.8-8.0 5.75 ml (2M)