Fusion rate depends on 3 main factors:

1- The electric pulse: Two DC pulses of 1.75 kV/cm for 15 micro seconds (BTX chamber is good for that).

2- The cell condition: trypsinzed cells with size less than 15 micro meter.

3- The fusion medium: 0.26 M mannitol + 0.1 mM MgSo4 + 0.5 mM HEPES + 0.05 BSA

Thanks! For the fusion medium, I use PVA instead of BSA.

with Hand made cloning you cand get the best fusion rate

You can get better fusion rates using needle electrodes

Thanks! I used needle electrodes to do pig cloning.