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From the analysis of the ratio of D peak intensity to G peak intensity in Raman spectroscopy, electron beam irradiation with energies of 100 eV was found to induce damage in single-layer graphene. The damage becomes larger with decreasing electron beam energy. Internal strain in graphene induced by damage under irradiation is further evaluated based on G peak shifts. The dose-dependent internal strain was approximately 2.22% cm2/mC at 100 eV and 2.65 × 10-2% cm2/mC at 500 eV. The strain induced by the irradiation showed strong dependence on electron energy.