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      • KCI등재

        Visual DNA Microarray Coupled with Multiplex-PCR for the Rapid Detection of Twelve Genetically Modified Maize

        Yongjin Li,Tao Xiong,Huawei Wu,Yazhen Yang 한국바이오칩학회 2016 BioChip Journal Vol.10 No.1

        We herein developed a visual DNA microarray system coupled with multiplex PCR (m-PCR) to rapidly detect twelve genetically modified maize (GMM). The microarray comprised short oligonucleotide probes complimentary to the specific gene region for twelve different GMM. The m-PCR products annealed to the microarray probe were reacted with streptavidin-alkaline phosphatase conjugate and nitro blue tetrazolium/5-bromo-4-chloro-3ʹ-indolylphosphate, p-toluidine salt (NBT/BCIP), resulting in blue spots that are easily visualized by unaided eyes for qualitative analysis. To ensure the reliability of this method, positive and negative hybridization controls were used in DNA microarray. Commercial GM materials (GMM: Bt176, Bt11, MON810, GA21, T25,MON88017, NK603, MON863, MON89034, DAS-59122-7, TC1507, MIR604; GM cotton: (MON1445, MON15985); GM soybean (Monsanto Roundup Ready soybean 40-3-2)) and non-GM materials were identified by this method and further confirmed by PCR and sequencing. The results showed that each probe consistently identified its corresponding GMM target very quickly and in a cost-effective and more time efficient way. The limit of detection is 0.5% for Bt176, Bt11, T25, MON88017, DAS59122-7, MON89034 and 1% for MON810, MIR604, GA21, MON863, NK603, TC1507. This method is advantageous because of rapid detection, cost-effectiveness and ease of use. These high specificity and sensitivity results demonstrate the feasibility of using this method in routine analysis of GMOs. We herein developed a visual DNA microarray system coupled with multiplex PCR (m-PCR) to rapidly detect twelve genetically modified maize (GMM). The microarray comprised short oligonucleotide probes complimentary to the specific gene region for twelve different GMM. The m-PCR products annealed to the microarray probe were reacted with streptavidin-alkaline phosphatase conjugate and nitro blue tetrazolium/5-bromo-4-chloro-3ʹ-indolylphosphate, p-toluidine salt (NBT/BCIP), resulting in blue spots that are easily visualized by unaided eyes for qualitative analysis. To ensure the reliability of this method, positive and negative hybridization controls were used in DNA microarray. Commercial GM materials (GMM: Bt176, Bt11, MON810, GA21, T25, MON88017, NK603, MON863, MON89034, DAS- 59122-7, TC1507, MIR604; GM cotton: (MON1445, MON15985); GM soybean (Monsanto Roundup Ready soybean 40-3-2)) and non-GM materials were identified by this method and further confirmed by PCR and sequencing. The results showed that each probe consistently identified its corresponding GMM target very quickly and in a cost-effective and more time efficient way. The limit of detection is 0.5% for Bt176, Bt11, T25, MON88017, DAS59122-7, MON89034 and 1% for MON810, MIR604, GA21, MON863, NK603, TC1507. This method is advantageous because of rapid detection, cost-effectiveness and ease of use. These high specificity and sensitivity results demonstrate the feasibility of using this method in routine analysis of GMOs.

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        A review of experimental methods for characterising composite viscosities of continuous fibre-reinforced polymer composites

        Wang Jinhuo,Han Yang,Ge Xiaohong,Qi Zhengbing,Zhao Jun,Wang Rongwen,Wu Huawei,Han Taiping,Sun Shaoxun,Wang Hui,Lin Jia,Liu Yuejun,Kong Xiangsong,Chen Qiming,Zeng Xiangxu 한국유변학회 2023 Korea-Australia rheology journal Vol.35 No.2

        Optimisation design of composite structures requires an accurate predictive model for forming behaviour. The simulation process contains a number of model parameters which include transverse and longitudinal viscosities of continuous fibrereinforced viscous composites, fundamental to predicting the shear rheology. Shearing the unidirectional composite along the fibre direction gives a measure of the longitudinal viscosity (LV), whilst shearing across or transverse to the fibre direction gives a measure of the transverse viscosity (TV). Numerous experimental work was conducted in the past to measure these two viscosities for various materials. However, conflicting measurements by different test methods were obtained and these apparent discrepancies had not yet been systematically investigated in any single study. This paper reviews previous work on characterisation techniques to further understand the cause of such discrepancy, and hence to improve measurement accuracy, which would benefit future work on theoretical modelling of the composite viscosities and optimisation simulation of composites forming. Some important findings, such as effects of resin-rich areas, contributory factors of elastic effects, non-Newtonian behaviour for composites with Newtonian matrix, aspect ratio and end effects of test samples, geometry effects of fibres and fibre rearrangement during shearing, existence of a mathematical relationship between LV and TV and necessary benchmarking exercise using Newtonian matrix composites, were summarised.

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