title: Event-specific detection of transgenic potato AV43-6-G7 using real-time and digital PCR methods.
abstract: 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.
  
  
  
  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 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.
  
  
  
  These results indicated that these event-specific methods would be useful for the identification of potato AV43-6-G7.
authors: Shandong Entry-Exit Inspection and Quarantine Bureau of People's Republic of China, Qingdao, China. ghw75@126.com.
authors: Hongwei
authors: H
authors: Gao
authors: Qingdao University Medical College, Qingdao, China.
authors: Xiaofan
authors: X
authors: Yu
authors: Chinese Academy of Inspection and Quarantine Institute, Beijing, China.
authors: Tingting
authors: T
authors: Deng
authors: Shandong Entry-Exit Inspection and Quarantine Bureau of People's Republic of China, Qingdao, China.
authors: Min
authors: M
authors: Sun
authors: Shandong Entry-Exit Inspection and Quarantine Bureau of People's Republic of China, Qingdao, China.
authors: Xizhi
authors: X
authors: Xiao
authors: Chinese Academy of Inspection and Quarantine Institute, Beijing, China.
authors: Xin
authors: X
authors: Huang
authors: Chinese Academy of Inspection and Quarantine Institute, Beijing, China.
authors: Ying
authors: Y
authors: Chen
authors: Qingdao University Medical College, Qingdao, China.
authors: Ronggui
authors: R
authors: Li
mesh_terms: D005504:Food Analysis / Q000379:methods*; D017343:Genes, Plant / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000145:classification / Q000235:genetics*; D011786:Quality Control; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D011198:Solanum tuberosum / Q000145:classification / Q000235:genetics*; D019076:Transgenes / Q000235:genetics*
keywords: Detection; Event Specific; Flanking sequence; Potato AV43-6-G7; Real-time PCR; Transgenic