Farmers in southern zone of Tigray are using different spacing below or above the national recommendation depending on the purpose of planting either for seed tuber or consumption due to lack of recommended plant spacing. This study was therefore conducted with the objective of determining the best inter and intra-row spacing for optimum tuber seed yield and quality of potato seed tuber at Ofla Wereda, Northern Ethiopia. Four different intra-row (20, 25, 30 and 35 cm) and inter-row (65, 70, 75 and 80 cm) spacing were used in the experiment. The result reveals that inter and intra-row spacing significantly (p<0.001) affected seed tuber yield ha-1 , the maximum seed tuber yield (36.89 and 37.54 ton ha-1) was recorded at 65 and 20 cm inter and intra-row spacing, respectively. From this study, it can be concluded that the narrow spacing (20 and 65 cm intra and inter-row spacing) produced higher seed tuber yield per hectare than other spacings. Thus, potato (Jalenie variety) growers in the study area can benefit if they use this narrow spacing (20 and 65 cm intra and inter-row spacing).