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Quantitative real-time PCR technique was used to determine the copy number of cry1Aa3 and cry1Ac in
transgenic chickpea lines. Gene specific primers designed to amplify relatively long amplicons (400 - 600
bp), were used to increase the specificity and sensitivity of real-time PCR using Syber green I as the
fluorescence indicator. Significantly accurate estimate of single transgene copy in the transformants was
achieved. Estimated copy number in transgenic lines using real-time quantitative PCR was found to be
same as analyzed by southern hybridization. The study shows the effectiveness of real-time PCR in
estimating transgene copy number in an efficient manner in transgenic chickpea plants.
Keywords: Real-time PCR, Chickpea transformation, Agrobacterium, copy number, Syber green.