Development of a standard, validated procedure for the isolation of transgene flanking regions in GM crops & detailed analysis of transgene insertion

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The isolation and analysis of transgene flanking regions forms a key component of molecular analysis and safety assessment of GM plants. A range of PCR-based techniques have been used for this purpose. During the G02 programme, these techniques were compared and the best methodology for cereals identified. This project will further develop the methodology and apply it to a large range of GM crops and different types of transgene insertion. New DNA walking kits will be compared to the existing technology to see if they offer advantages in throughput, reproducibility and cost. The project will provide standard, validated methods for the analysis of junction sequences in a range of GM crops, containing transgene insertions of different complexity. The methods developed will be validated by the analysis of a standard set of GM samples at two different sites. In selected lines, the entire transgene insertion site will be sequenced in addition to the transgene flanking regions. The project will provide valuable information on the patterns of transgene insertion in a range of GM crops as well as robust methodologies for use in GM crop safety assessment.

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