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Measurement and simulation studies are conducted on transmission crosstalk in thick silicon-on-insulator substrates. This paper focuses on the role of buried oxide layers and deep trench isolation in suppressing crosstalk. With radio frequency coupling paths on substrates depending on noise frequency, a deep trench guides noise signals to substrates and suppresses transmission crosstalk between input and output ports. Good agreement is obtained between electromagnetic field (EM) simulation and measurement results. The EM simulation results suggest different approaches might be used in designing deep trench isolation patterns for a middle resistivity (MR) substrate and a high resistivity (HR) substrate. Deep trench isolation plays a more important role in HR substrates than in MR substrates.