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The quality, structural defects, and transport properties of GO can be tailored by maintaining ultrasonic bath temperature. Herein, the effect of temperature-controlled exfoliation on the structural defects and transport properties of the resulting graphene oxide (GO) sheets are examined. The large crystallite size (12.23 nm) and low defect density (2.16 × 10 ^11 cm ^−2 ) are found at 20 °C exfoliations. Highly oriented GO sheets were exfoliated at 20 °C which was confirmed through Bernal-stacking proportions (∼30%). Furthermore, electrical transport properties are high at 10 °C exfoliation temperature while optimum at other temperatures, calculated through impedance spectroscopy. Comparatively, smooth, wrinkle-free, and lesser bulges are found at 20 °C and 30 °C exfoliation temperatures.