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The authors investigated the influence of substrates on Raman scattering spectrum from graphene. The room-temperature Raman signatures from graphene layers on GaAs, sapphire, and glass substrates were compared with those from graphene on the standard Si∕ Si O 2 (300 nm) substrate, which served as a reference. It was found that while G peak of graphene on Si∕ Si O 2 and GaAs is positioned at 1580 cm− 1⁠, it is downshifted by∼ 5 cm− 1 for graphene on sapphire and, in some cases, splits into doublets for graphene on glass with the central frequency around 1580 cm− 1⁠. The obtained results are important for nanometrology of graphene and graphene-based devices.