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  • Title: [A quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR)].
    Author: Zhang B, Huang JQ, Wei ZM.
    Journal: Shi Yan Sheng Wu Xue Bao; 1999 Jun; 32(2):207-11. PubMed ID: 12548787.
    Abstract:
    In order to obtain single T-DNA copy transgenic rice, we have established a quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR). IPCR was used to amplify junction fragments, i.e. plant genomic DNA sequences flanking the known T-DNA sequences, which will help to estimate the T-DNA copy number in transgenic rice. We have analyzed 20 transgenic plants of 15 transgenic lines. Most plants (12) contain one integrated T-DNA copy per genome, 3 plants contain two and 1 plant contains 3 copies. In 4 transgenic plants no T-DNA copies could be detected using this method. The IPCR results were further tested by Southern analysis and sequence analysis.
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