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A Combined Protocol for PCR Detection of GM in Seed - VS0136

This work involves the development, optimisation and assessment of a combined PCR based approach to seed testing which can provide both % GM seed and % GM DNA results in order to improve the reliability of borderline test decisions and provide results to inform decisions for seed and crop thresholds, which may be expressed differently in legislation. The required development and testing of the protocol will also provide accurate estimates of uncertainty associated with different thresholds and measured % GMO results.
1. Obtain GM materials for use as standard genotypes in protocol testing with known variable copy number and genotype. Establish reliable PCR tests for these lines.

2. Make mixtures of seed with variable % GM seed and known % GM DNA. Theoretically examine the statistics behind optimisation of seed bulk size and number with respect to accuracy and cost.

3. Test seed mixtures for expected binomial distribution of positive tests in stage 1 of protocol. Statistically test results and establish best size and number of Stage 1 bulks.

4. Optimise stage 2 real-time real-time PCR using seed bulks of known GM composition.

5. Combined method testing. Using materials and optimised PCRs from objectives 1-4, replicate the entire procedure several times to measure accuracy of accept/reject decisions. Compare results statistically to expected values.
Project Documents
• Final Report : A Combined Protocol for PCR Detection of GM in Seed   (279k)
• Final Report - Annex : A Combined Protocol for PCR Detection of GM in Seed   (160k)
Time-Scale and Cost
From: 2005

To: 2006

Cost: £49,346
Contractor / Funded Organisations
Central Science Laboratory
Plant Varieties and Seeds              
Plants and Animals              
Sustainable Farming and Food              
Sustainable Production              
Fields of Study
Plant Varieties and Seeds