| Info. | Vol.10 - No.1 (2016.03.20) |
|---|---|
| Title | Visual DNA Microarray Coupled with Multiplex-PCR for the Rapid Detection of Twelve Genetically Modified Maize |
| Authors | Yongjin Li1,2,*, Tao Xiong2, Huawei Wu2 & Yazhen Yang2 |
| Institutions | 1College of Life Sciences, Huzhou University, Huzhou 313000, China 2College of Life Sciences, Yangtze University, Jingzhou 434025, China |
| Abstract | 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-3賈-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. |
| Keyword | Visual detection, Multiplex PCR, Genetically modified maize, Microarray, Biochip |
| PDF File |
 # 2010년도 발행분 부터는 Springer 의 BioChip Journal 페이지에서 전문을 열람하실 수 있습니다.
# 학회회원 로그인 후 [ Springer BioChip Journal 열람하기 ] 버튼을 클릭하시면 새창으로 열립니다. |
