All Rights Reserved AEIC 2004 0 1 2 3 Response 0.10.512 Concentration 5% CV 20% CV Error bars for concentrations determined by the.

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All Rights Reserved AEIC 2004

Response Concentration 5% CV 20% CV Error bars for concentrations determined by the method having a 20% CV overlap. Both methods give the same concentration but it is not possible to say that adjacent concentrations differ from each other in the assay with the 20% CV within the given confidence interval.

All Rights Reserved AEIC 2004

y = x R 2 = % Amount of analyte Optical Density LOD LOQ Quantitative Range Linear regression Correlation coefficient All Rights Reserved AEIC 2004

% GMO Corn OD Event176 Bt11 Mon810

All Rights Reserved AEIC 2004 y = x x R = %GMO OD 0.1% LOD Soymeal was toasted for 60 min. at 100 ºC Antibody not selective for toasted soymeal Antibody detects toasted soymeal

All Rights Reserved AEIC 2004

During routine analysis:  Follow strict rules how to work cleanly in a molecular biology lab  Use one way sample flow lab design to prevent contamination with positive DNA from down- stream steps (especially PCR products), strict separation of certain lab areas from each other  Use duplicate analysis of each sample  Use negative controls on homogenization, DNA extraction and PCR setup Corruption of sample, extracted sample DNA or PCR setup contains positive DNA During validation:  Search data base for potential unwanted (homologous) annealing sites  Test new primer system on a wide range of DNA types that appear in food samples  Verify the identity of the PCR product by Southern blot or restriction enzyme digestion Artifact PCR products from templates other than the target sequence in question (worst scenario: artifacts of the same size as the expected PCR product) Control MeasuresProblem All Rights Reserved AEIC 2004

During validation:  Test primer system on sample DNA solutions known to contain inhibitory compounds During routine analysis:  Test each PCR reaction for inhibition by an individual positive control (a spiked counterpart)  Use duplicate analysis of each sample Inhibition of PCR reaction During design and validation:  Choose appropriate extraction protocol for sample type  Run controls to monitor efficiency of each extraction series DNA extraction failed During design and validation:  Use computer aided primer design  Test the sensitivity of new primer systems for the intended concentration range Insufficient sensitivity of primer system Control MeasuresProblem All Rights Reserved AEIC 2004