Statistical tools for transgene copy number estimation based on real-time PCR

Background: As compared with traditional transgene copy number detection technologies such as Southern blot analysis, real-time PCR provides a fast, inexpensive and high-throughput alternative. However, the real-time PCR based transgene copy number estimation tends to be ambiguous and subjective stemming from the lack of proper statistical analysis and data quality control to render a reliable estimation of copy number with a prediction value. Despite the recent progresses in statistical analysis of real-time PCR, few publications have integrated these advancements in realtime PCR based transgene copy number determination. Results: Three experimental designs and four data quality control integrated statistical models are presented. For the first method, external calibration curves are established for the transgene based on serially-diluted templates. The Ct number from a control transgenic event and putative transgenic event are compared to derive the transgene copy number or zygosity...

• Type: Journal
• Year: 2007
• Where: BMCBI
• Authors: Joshua S. Yuan, Jason N. Burris, Nathan R. Stewart, Ayalew Mentewab, C. Neal Stewart Jr.
• Keywords: BMCBI 2007, Copy Number, Transgene, Transgene Copy Number