Primer and probe for real-time fluorescent quantitative PCR detection of transgenic alfalfa J101 strain Taqman and detection method

• Main Authors: LYU YINGZI, WANG DINGGUO, YAO BAIHUI, JIANG XIANG, LIN HUIJIAO, ZHENG GAOBIN, LIU ERLONG, TANG JIE, QIN ZHUOMIN, FAN WUJIANG, LU LI, LIN XUEQIN
• Format: Patent
• Language: English
• Published: 29.04.2015
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a primer and a probe for real-time fluorescent quantitative PCR detection of a transgenic alfalfa J101 strain Taqman and a detection method. For the transgenic alfalfa strain J101 not approved by Chinese Department of Agriculture with an agricultural transgenic organism safety certificate, the primer and the probe are designed according to the adjacent area sequence of the end 5', and the specific real-time fluorescent quantitative PCR detection method for the transgenic alfalfa J101 strain is established successfully. Results indicate that the standard curve of the established specific real-time fluorescent quantitative PCR detection method for the J101 strain is excellent in correlation coefficient, amplification efficiency and repeatability, the minimum detection DNA concentration is 16pg, which is equivalent to 10 copies of transgenic alfalfa strain J101 genome DNAs, and the minimum quantitative detection DNA concentration is 80pg, which is equivalent to 50 copies of J101 genome DNAs; and as a result, the detection method can be applied to qualitative and/or quantitative detection of the J101 strain quickly, accurately and stably.