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Development of Detection and Quantification Methods for Transgenic Potato Using Multiplex PCR and Plasmidic Reference Material
Primers and probes used in PCR, mPCR, recombinant PCR, and real-time PCR (RT-qPCR) in this research.
Primers and probes used in PCR, mPCR, recombinant PCR, and real-time PCR (RT-qPCR) in this research.

Author:Yuan-Kai Tu, Yu-Wei Feng, Min-Tze Wu, Shi-Yu Wang, Shu Chen, and Han-Wei Chen*

Abstract:

    The objectives of this research were to establish a new quantification method and a fast multitarget PCR method for genetically modified potato event, EH92-527-1. A plasmidic reference material (PRM), pGEM-EH92, was constructed and used as a calibrant to quantify content of EH92-527-1. Results showed the lowest of quantification was 0.1% (w/w), bias and relative standard error were 16.3% and 6–12.9%, respectively, at 3% (w/w) true value. It indicated that the quantification method by means of PRM was able to practically determine the EH92-527-1 event content of mixed sample above 3%. When RNApol (RNA polymerase), nptII (Neomycin phosphotransferase II) and antisense GBSS (Granule bound starch synthase) sequences of EH92-527-1 event were used as multiplex PCR target, the lowest of qualification of multiplex PCR method was 3%.

Key words:Genetically modified, Potato, Real-time PCR, Plasmidic reference material

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Journal of
Taiwan Agricultural Research
Vol. 73, No.3 (2024) ISSN: 2790-086X Vol. 73, No.3 (2024)
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