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Validation of the Method for Event Specific Mon810 and NK603 and Endogenous Gene Identification by using Multiplex Real-time PCR Technique |
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| รหัสดีโอไอ | |
| Creator | Piyanuch Sornchai |
| Title | Validation of the Method for Event Specific Mon810 and NK603 and Endogenous Gene Identification by using Multiplex Real-time PCR Technique |
| Contributor | Nattawadee Buntongdee, Thitirut Assawamongkolsiri, Khanitha Wongwathanarat |
| Publisher | Department of Agriculture |
| Publication Year | 2562 |
| Journal Title | Thai Agricultural Research Journal |
| Journal Vol. | 37 |
| Journal No. | 3 |
| Page no. | 224-237 |
| Keyword | genetically modified maize, multiplex real-time PCR technique, Mon810, NK603, validation |
| URL Website | http://at.doa.go.th/journal |
| Website title | Thai Agricultural Research Journal |
| ISSN | 0125-8389 |
| Abstract | Genetically modified maize has been widely accepted for commercialization. The most popular approved events are lepidopteran insect resistance (Mon810) and glyphosate herbicide tolerance (NK603). However, importation conditions of GM products insome countries requires identification of the GM event and biosafety assessment.At present, the international laboratory analyzes GM event by using simplex real-time PCRtechnique which is lengthy, contains many steps and is prone to contamination.The multiplex real-time PCR had been developed for detecting multiple modified genesat the same time. We validated this method for GM maize: Mon810 and NK603 eventsdetection. The results revealed that Mon810, NK603 and HMG primers and probes wereabsolutely specific to Mon810 and NK603 maize. The parameters of this multiplexreal-time PCR method were within the acceptable parameter standard including 97 to112 percentage of PCR efficiency, -3.38 to -3.07 of slope, 0.99 of linearity (R2), limit ofdetection in 0.1 percentage and no false positive or negative results. In summary, multiplexreal-time PCR in this study can be used for the events screening method of GM maizeand maize products |
