中文摘要：本发明提供了引物对和探针组合，本发明还提供了所述的引物对和探针组合在检测或辅助检测转基因大豆MON87701和转基因大豆MON87708中的应用；本发明同时还提供了含有该引物对和探针组合的试剂盒及应用，以及基于引物对和探针组合或试剂盒的检测转基因大豆MON87701和转基因大豆MON87708的方法。经实验证明，本发明的引物对和探针组合特异性好、灵敏度高、检测效率高，且基于该引物对和探针组合建立的转基因大豆MON87701和转基因大豆MON87708的双重荧光PCR方法高通量、灵敏、准确和快速，为同时检测转基因大豆MON87701和转基因大豆MON87708提供了更有效的方法。
 
外文摘要：NOVELTY - A primer pair and probe combination comprising (a) primer pairs comprising base pair sequences of SEQ ID NOs: 1 and 2, and probe comprising base pair sequence of SEQ ID NO: 3, and (b) set of primer pairs comprising base pair sequences of SEQ ID NOs: 4 and 5, and probe comprising base pair sequence of SEQ ID NO: 6, is new.
USE - The primer pair and probe combination is useful in kit and preparation of product for performing detection or assisted detection of transgenic soybean MON87701 and transgenic soybean MON87708 (all claimed).
ADVANTAGE - The primer pair and probe combination detects transgenic soybean rapidly and accurately with good specificity, high sensitivity, high detecting efficiency, and high throughput, and enables to simultaneously detect transgenic soybean MON87701 and transgenic soybean MON87708.
DETAILED DESCRIPTION - A primer pair and probe combination comprising (a) primer pairs comprising base pair sequences of SEQ ID NOs: 1 and 2, and probe comprising base pair sequence of SEQ ID NO: 3, and (b) set of primer pairs comprising base pair sequences of SEQ ID NOs: 4 and 5, and probe comprising base pair sequence of SEQ ID NO: 6, is new. atttcccggacatgaagcc (SEQ ID NO: 1), tggttagtgagtggcagtaatcg (SEQ ID NO: 2), caagctaattcaagaaagtgaaggcacg (SEQ ID NO: 3), gcttatccgatttgagcattg (SEQ ID NO: 4), cgtttcccgccttcagttt (SEQ ID NO: 5) and atgagccatttagtctcaccttcaaaca (SEQ ID NO: 6). INDEPENDENT CLAIMS are included for the following:
(1) kit comprising the primer pair and probe combination; and
(2) method for performing detection or assisted detection of transgenic soybean MON87701 and transgenic soybean MON87708, involves using the primer pair and probe combination or the kit.
TF   TECHNOLOGY FOCUS - BIOTECHNOLOGY - Preferred Kit: The kit further comprises a fluorescent quantitative PCR amplification reagent comprising deoxynucleoside triphosphate, magnesium ion, PCR reaction buffer liquid and Thermus aquaticus (Taq) DNA polymerase. Preferred Method: The method involves performing dual fluorescent PCR amplification of the biological sample containing DNA as template and the primer pair and probe combination, and determining whether the biological sample is transgenic soybean MON87701 or transgenic soybean MON87708 based on the results of PCR amplification or whether the biological sample contains the strain specific gene component of transgenic soybean MON87701 and/or transgenic soybean MON87708. The method involves performing dual fluorescent PCR amplification using a reaction system comprising 10 mu l Probes Master 2x conc, 0.4-0.6 mu l 10 mu mol/l primer pair, 0.05-0.3 mu l 10 mu mol/l probe, 1-2 mu l 100-300 ng/ mu l DNA template, and up to 20 mu l double distilled water. The PCR amplification reaction is performed by pre-denaturing at 95 degrees C for 3 minutes, denaturing at 95 degrees C for 5-10 seconds, preferably 5 seconds, annealing at 59 degrees C for 25-40 seconds, preferably 30 seconds, and repeating for 40 cycles.
 
主权项：引物对和探针组合，其特征在于，其中一组引物对包含SEQ?ID?NO：1和SEQ?ID?NO：2所示的核苷酸序列，与这组引物对相对应的探针包含SEQ?ID?NO：3所示的核苷酸序列；另一组引物对包含SEQ?ID?NO：4和SEQ?ID?NO：5所示的核苷酸序列，与这组引物对相对应的探针包含SEQ?ID?NO：6所示的核苷酸序列。
 
申请号：CN201610486875.6
专利号：CN106399302-A
公开/公告号：CN106399302-A
申请/专利权人：汕头出入境检验检疫局检验检疫技术中心；伊犁出入境检验检疫局综合技术服务中心；中国检验检疫科学研究院
发明/设计人：段建发;周广彪;乾义柯;魏霜;付伟;许如苏;温尔英;陈文婉
分类号：C12N15/11;C12Q1/68
主分类号：C12N15/11
法律状态：实质审查
点击下载：一种同时高效检测转基因大豆MON87701和MON87708品系特异性基因的双重荧光PCR方法