This study is a descriptive research aimed at accessing and describing customers’ expectation and perception of service quality in the banking industry of Ghana, and subsequently identifying the dimensions of service quality that drives customer perceived service quality in the banking industry of Ghana. The target population of the study was customers of universal banks in Ghana. In all 1000 customers of universal banks in Ashanti region were selected as the sample size of the study. The findings of the study revealed that the critical factors of service quality among universal banks in Ghana are five factors which are named: Physical Environment, Reliability, Responsive, Assurance and Empathy. Customers’ perception of the level of excellence of service quality of the universal banks was found to be below their expectation. However, customers’ perception of the level of excellence of physical environment dimension and the reliability dimension were found to be of customers’ expectations. The findings of the study revealed that the most influential dimension that drives customers perception of overall service quality is the empathy dimension. It was found that background characteristics such as gender, age, income level, and academic qualifications of customers of universal banks have significant influence on customers’ perception and expectation of service quality. It is recommended that greater emphasis should be placed on empathy since it is found to be the most influential factor of overall service quality perception of customers of universal banks in Ghana. Policy makers of universal banks should factor in customers’ background characteristics when formulating policies on improving service quality. Future studies are encouraged to perform a cross country comparison and compare their findings in other African countries with the results of current study.