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)

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