Detection of Genetically Modified Ingredients in Transgenic Papaya using Universal Primer-multiplex PCR
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Abstract:
In order to meet the need for simultaneous detection of multiple targets and to overcome the several limitations associated with traditional multiplex polymerase chain reaction (PCR) such as primer exclusion, poor reproducibility etc., a novel universal primer-multiplex PCR approach was developed to efficiently and rapidly detect the genetically modified ingredients in papaya. Fresh papaya leaves from three transgenic papaya cultivars (Event 55-1, “GM - YK”, “Huanong No. 1”) and non-transgenic papaya (“Tainong No. 2”) were chosen for this study. Five specific primers were designed based on the sequences of the exogenous genes of the three transgenic papaya cultivars and endogenous reference gene (papaya proteinase). The electrophoretic bands of the composite primers (with universal adapter primers) were brighter and clearer in both simplex PCR and multiplex PCR detections, compared to the PCR results of primers without the addition of the adapters. The sensitivity of multiplex PCR using universal primers was one order of magnitude higher than that detected using common primers. Compared to ordinary multiplex PCR, the novel universal primer-multiplex PCR has a lower detection limit and higher sensitivity in the detection of genetically modified ingredients in transgenic papaya, providing a new technique for the rapid detection of genetically modified ingredients.
