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Plant Molecular Biology Reporter

, Volume 21, Issue 1, pp 73–80 | Cite as

Rapid PCR-based determination of transgene copy number in rice

  • Fengling Li
  • Moul Dey
  • Chengkun He
  • Vinod Sangwan
  • Xianzhong Wu
  • Ray Wu
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Abstract

We present a simple, rapid, and low-cost method to determine transgene copy number in rice. More than 100 first- and second-generation transgenic rice plants were tested. The plasmid (pRCopy) used for rice transformation contains the specific gene of interest and a partially deleted cytochrome c gene (cyc), a single-copy gene in rice. A 132-bp segment of the cloned ricecyc was shortened to 108 bp by deleting a 24-bp internal fragment. After PCR amplification of the genomic DNA from transgenic rice harboring pRCopy, the 2 expected bands were found. The 121-bp band corresponds to the endogenouscyc; the 97-bp band comes from the integrated pRCopy. Clear distinctions can be made between single and multiple copies of the transgene by comparing band densities.

Key words

rapid PCR method transgene copy number 

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Copyright information

© International Society for Plant Molecular Biology 2003

Authors and Affiliations

  • Fengling Li
    • 1
  • Moul Dey
    • 1
  • Chengkun He
    • 1
  • Vinod Sangwan
    • 1
  • Xianzhong Wu
    • 1
  • Ray Wu
    • 1
  1. 1.Department of Molecular Biology & GeneticsCornell UniversityIthacaUSA

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