A novel quadruplex real-time PCR method for simultaneous detection of Cry2Aeand two genetically modified cotton events (GHB119 and T304-40)

Li, Xiang; Wang, Xiuxiu; Yang, Litao; Liu, Yueming; He, Yihua; Pan, Liangwen

doi:10.1186/1472-6750-14-43

Cited by 8 publications

(10 citation statements)

References 11 publications

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“…As expected, following the event-specific qualitative PCR assay of AV43-6-G7 potato, a passive real-time PCR amplification curve was obtained for AV43-6-G7 potato, whereas no amplification curve was observed for the other GM lines [26] (Fig. 3).…”

Section: Resultssupporting

confidence: 72%

Event-specific detection of transgenic potato AV43-6-G7 using real-time and digital PCR methods

Gao

Deng

et al. 2016

BMC Biotechnol

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BackgroundThe isolation of unknown DNA sequences flanked by known sequences is an important task in the event-specific detection of GMOs. None of event-specific detection method was developed based on the junction sequence of an exogenous integrant in the transgenic potato AV43-6-G7.ResultsThe flanking sequence between the exogenous fragment and recombinant chromosome of this potato was successfully acquired through exogenous gene 5′-RACE. The event-specific primers and Taqman probe were designed to amplify fragments spanning the exogenous DNA and potato genomic DNA. The specific real-time PCR and digital PCR detection methods for AV43-6-G7 potato were established based on primers designed according to the flanking sequences. The detection limit of the qualitative PCR assay was 0.01 % for AV43-6-G7 potato in 100 ng of potato genomic DNA, corresponding to approximately 11.6 copies of the potato haploid genome. The ddPCR assays for Potato AV43-6-G7 achieved a limit of quantification of approximately 58 target copies, with RSD ≤ 25 %. The aLOQ of this system was approximately 1.2 copies.ConclusionsThese results indicated that these event-specific methods would be useful for the identification of potato AV43-6-G7.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-016-0303-8) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 72%

Event-specific detection of transgenic potato AV43-6-G7 using real-time and digital PCR methods

Gao

Deng

et al. 2016

BMC Biotechnol

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“…In Fig. 1, the RSD values of the eight genes were from 0.93 to 4.54%, which were all in the acceptable range [16]. Especially, the RSD values of ClACT, ClEF1a, and ClUBC2 were less than 1%.…”

Section: Gene Expression Analysismentioning

confidence: 87%

Abiotic stress and tissue-specific reference genes for quantitative reverse transcription PCR analysis in Korean native watermelons, Citrullus lanatus ‘Black-King’ and ‘Speed-Plus-Honey’

et al. 2018

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A wide variety of research on watermelon has been conducted, and such studies have been motivated by the published genome sequence database of watermelon. Screening of proper reference genes is the primary step for normalization in gene expression analysis. Based on previous studies conducted on Arabidopsis and cucumber, we selected eight candidate reference genes of ClACT, ClEF1a, ClGAPDH, ClIDH, ClLUG, ClPTB, ClUBC2, and Cl18SrRNA, respectively, encoding b-Actin, elongation factor 1-a, glyceraldehyde-3-phosphate-dehydrogenase, NADP-isocitrate dehydrogenase, leunig, polypyrimidine tract-binding protein1, ubiquitin-conjugating enzyme E2, and 18S ribosomal RNA from watermelon (Citrullus lanatus). The expression levels of these eight genes were evaluated by RT-qPCR under plant hormone-treatment (100 lM ABA) and abiotic stresses such as drought, cold (4°C), and high salt concentration (250 mM NaCl). The expression patterns of these eight genes were further compared across different types of watermelon tissues such as flower, leaf, tendril, stem, root, and whole seedling. Our results showed that expressions of ClACT and ClEF1a, respectively in the Korean native watermelon cultivars Citrullus lanatus 'Black-King' and 'Speed-Plus-Honey' were least affected by the environmental stresses regardless of tissue types. Here, we suggest two ideal reference genes for watermelon RT-qPCR-based gene expression study.

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“…Capture probes hybridize with homologous amplified target DNA labeled with a fluorescent dye or a chemiluminescence dye, which can be detected by the naked eye or a laser scanner. Due to the limited stability and repeatability of RDB methods, RDB is unsuitable for routine analysis of GMC events in imported and exported goods [3]. This capacity is much greater than other DNA detection methods, such as PCR and LAMP.…”

Section: Reverse Dot Blot (Rdb) Methodsmentioning

confidence: 99%

“…According to the International Service for the Acquisition of Agribiotech Applications, GMC crops have been commercially cultivated for over 17 years, and the global planting area has increased 100-fold [1][2][3]. Along with the rapid development of GMCs and increasing numbers in food, feed, and the environment, GMCs have come under suspicion from governments and citizens because of potential safety risks [4].…”

Section: Development Of Gmcsmentioning

confidence: 99%

Detection of genetically modified components of rice using a membrane-based array method

Zhang¹,

Qiu²,

Huang³

et al. 2016

Electronics, Electrical Engineering and Information Science

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A method for the detection of genetically modified components (GMCs) is reported for detection of GMCs in rice plants and foods using a membrane-based array. A DNA sequence as probes was immobilized on modified paper film-chips to form the sensing array surface. The target DNA from sample was amplified by polymerase chain reaction (PCR) using labeled biotin primers. The PCR products were added to the sensing surface using an automatic membrane-based array instrument (MAI). When the target DNA passed the surface, it was captured by the complementary probes. The signal was visualized with a streptavidin/alkaline phosphatase (AP) and AP color development kit. Compared to manual dot blotting, the MAI could automatically and rapidly finish several GMC types of rice detection, such as BT11, TC1507, Mon88017, and Mon89034. Stable 1 Sheng-Sheng Zhang 2 Li-Chun Qiu Electronics, Electrical Engineering and Information Science Downloaded from www.worldscientific.com by LA TROBE UNIVERSITY on 06/16/16. For personal use only. and consistent qualitative results were acquired. The sensitivity reached 0.1%. Thus, the automatic MAI might be a valuable GMC detection tool.

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A novel quadruplex real-time PCR method for simultaneous detection of Cry2Aeand two genetically modified cotton events (GHB119 and T304-40)

Cited by 8 publications

References 11 publications

Event-specific detection of transgenic potato AV43-6-G7 using real-time and digital PCR methods

Event-specific detection of transgenic potato AV43-6-G7 using real-time and digital PCR methods

Abiotic stress and tissue-specific reference genes for quantitative reverse transcription PCR analysis in Korean native watermelons, Citrullus lanatus ‘Black-King’ and ‘Speed-Plus-Honey’

Detection of genetically modified components of rice using a membrane-based array method

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