Event-specific detection of transgenic potato AV43-6-G7 using real-time and digital PCR methods. Issue 1 (December 2016)

• Record Type: Journal Article
• Authors: Gao, Hongwei; Yu, Xiaofan; Deng, Tingting; Sun, Min; Xiao, Xizhi; Huang, Xin; Chen, Ying; Li, Ronggui
• Is Part Of: BMC biotechnology. Volume 16:Issue 1(2016)
• Page Start: 1
• Page End: 9
• Publication Date: 2016-12
• Subjects: Transgenic -- Potato AV43-6-G7 -- Flanking sequence -- Event Specific -- Real-time PCR -- Detection; Biotechnology -- Periodicals; 660.605
• DOI: 10.1186/s12896-016-0303-8
• Languages: English
• ISSNs: 1472-6750

Abstract

Abstract Background The 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. Results The flanking sequence between the exogenous fragment and recombinant chromosome of this potato was successfully acquired through exogenous gene 5′-RACE. The event-specific primers andTaq man 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. Conclusions These results indicated that these event-specific methods would be useful for the identification of potato AV43-6-G7.