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The development of efficient crop transformation systems has necessitated the development of efficient methods for detailed molecular characterization of putative events. This chapter details the routine use of quantitative real-time polymerase chain reaction to determine transgene copy number in putative transgenic events. This approach has allowed the analysis of plantlets in tissue culture prior to transfer to soil and greenhouse. Implementation of the TaqMan transgene copy assay permits the efficient utilization of limited resources and space to develop a highly efficient transgenic event production pipeline. Other applications for this assay within the biotechnology production pipeline are also discussed.