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Development Triplex Real-Time PCR Screening Method to Detect GM Maize for ISO/IEC17025 Standard Accreditation |
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| รหัสดีโอไอ | |
| Creator | 1. Piyarat Thammakijjawat 2. Piyanuch Sornchai 3. Thitirut Assawamongkolsiri 4. Pongsakorn Sanvittayakul |
| Title | Development Triplex Real-Time PCR Screening Method to Detect GM Maize for ISO/IEC17025 Standard Accreditation |
| Publisher | Department of Agriculture |
| Publication Year | 2562 |
| Journal Title | Thai Agricultural Research Journal |
| Journal Vol. | 36 |
| Journal No. | 3 |
| Page no. | 316-331 |
| Keyword | Genetically Modified Corn, Screening Method, Multiplex Real Time PCR, Triplex Real Time PCR ข้าวโพดดัดแปลงพันธุกรรม, การตรวจคัดกรองพืชดัดแปลงพันธุกรรม, CaMV35S Promoter, Nos terminator, hmg gne, Triplex Real-time PCR |
| URL Website | http://at.doa.go.th/journal |
| Website title | Thai Agricultural Research Journal |
| ISSN | 0125-8389 |
| Abstract | Analysis of Genetically Modified plants in general laboratory uses qualitative PCR todetect CaMV 35S promoter and NOS terminator by single Real-Time PCR method. Thequality of the DNA test was examined using plant specific reference gene, in the case ofmaize, the hmg gene was tested by PCR or Real-time PCR. This research has developeda method for screening genetically modified maize by testing three target genessimultaneously in a single reaction. The Triplex Real-Time PCR assay of CaMV 35Spromoter, NOS terminator, and maize hmg gene were tested in the same reaction. Resultsshow that the developed screening method was specific and sensitive for certified referencematerials of corn-GM events. The Limit of detection was found at 0.01%. The concentrationof the DNA template is 50-200 nanograms per reaction. Seventeen maize samples were alsotested with this newly developed method. The test results were comparable to those ofSingle plex Real time PCR but this method was more efficient because it reduced bothcost and time for the analysis It can be used as a standard screening method for maizeand maize products. The method has already been approved as ISO / IEC17025 standardfrom Bureau of Laboratory Standards and used as a standard test method in laboratory ofDepartment of Agriculture. |
