[Co1.0/Pt0.75]n multilayers consisting of 1.0-nm-thick Co and 0.75-nm-thick Pt layers with n = 2–6 were grown on glass substrates with and without a Pt underlayer at room temperature (RT). In this study, the effects of total thickness and Pt underlayer on magnetic properties and microstructures are investigated. Without a Pt underlayer, the [Co1.0/Pt0.75]n film exhibits soft magnetic behaviors due to poor crystallization. After the inclusion of a Pt underlayer between magnetic layer and glass substrate, the [Co1.0/Pt0.75]n films show significant perpendicular magnetic anisotropy (PMA), out-of-plane coercivity, and squareness as the total thickness of [Co1.0/Pt0.75]n is in the range of 5.25–10.5 nm. The induced PMA is attributed to the epitaxial growth of CoPt grains along Pt(111) underlayer. The RT-prepared CoPt film with PMA shows high potential in future applications of spintronics.