The positive and negative charging difference in the contact charge electrophoresis of a water droplet suspended in oil was investigated to find out the origin of this charging difference. Through numerous experiments and numerical analysis, the charging difference has been found to be mainly originated from electrostatic sources. Two electrostatic sources were found in the present experimental setup, and by excluding those two sources the charging difference was successfully diminished. The present findings well explain previous experimental results and also provide design guidelines for consistent droplet movement control in contact charge electrophoresis-based digital microfluidic systems. Finally, further discussions on the obtained results, its implications, and future work are discussed.