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Plant Cell Reports

, Volume 20, Issue 10, pp 948–954 | Cite as

Quantitative real-time PCR as a screening tool for estimating transgene copy number in WHISKERS™-derived transgenic maize

  •  P. Song
  •  C. Cai
  •  M. Skokut
  •  B. Kosegi
  •  J. Petolino
Genetic Transformation and Hybridization

Abstract.

Quantitative real-time PCR (qRT-PCR) was adapted to estimate transgene copy number in transgenic maize callus and plants. WHISKERS™-derived transgenic callus lines and plants were generated using two different gene constructs. These transgenic materials represented a range of copy number. A 'standard curve' was established by mixing plasmid DNA with non-transgenic genomic maize DNA using a calculated ratio of target gene to host genome size. 'Estimated' copy number in the callus lines and plants using qRT-PCR was correlated with the 'actual' copy number based on Southern blot analysis. The results indicated that there was a significant correlation between the two methods with both gene constructs. Thus, qRT-PCR represents an efficient means of estimating copy number in transgenic maize.

Quantitative real-time polymerase chain reaction Transgene copy number Maize transformation WHISKERS™ transformation Transgene insertion complexity 

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Copyright information

© Springer-Verlag 2002

Authors and Affiliations

  •  P. Song
    • 1
  •  C. Cai
    • 1
  •  M. Skokut
    • 1
  •  B. Kosegi
    • 1
  •  J. Petolino
    • 1
  1. 1.Dow AgroSciences LLC, 9330 Zionsville Rd., Indianapolis, IN 46268, USA

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