In this study, a novel universal primer-multiplex-PCR (UP-M-PCR) method adding a universal primer (UP) in the multiplex
PCR reaction system was described. A universal adapter was designed in the 59-end of each specific primer pairs which
matched with the specific DNA sequences for each template and also used as the universal primer (UP). PCR products were
analyzed on sequencing gel electrophoresis (SGE) which had the advantage of exhibiting extraordinary resolution. This
method overcame the disadvantages rooted deeply in conventional multiplex PCR such as complex manipulation, lower
sensitivity, self-inhibition and amplification disparity resulting from different primers, and it got a high specificity and had a
low detection limit of 0.1 ng for single kind of crops when screening the presence of genetically modified (GM) crops in
mixture samples. The novel developed multiplex PCR assay with sequencing gel electrophoresis analysis will be useful in
many fields, such as verifying the GM status of a sample irrespective of the crop and GM trait and so on.