中文摘要：一种用于转基因玉米多重检测的微滴PCR偶联DGGE分析方法，首先，针对待测样品分别设计PCR特异性引物对，进行微滴PCR扩增，扩增产物经纯化后进行DGGE电泳，并进行SYBRGreen染色，在紫外光下显影观察，来检测转基因玉米基因组的多重DNA分子。该方法可以实现高通量分析多个DNA目标核酸分子，且灵敏度高，内标基因的稳定性也很好。因此，本发明利用微滴PCR偶联DGGE技术实现了生物基因定性、高通量检测。
外文摘要：NOVELTY - A primer set for qualitative and high-throughput multiplex detection of transgenic corn comprising specific primer pairs for transgenic corn BT176, TC1507, and MON863, is new.    USE - The primer set is useful for qualitative and high-throughput multiplex detection of transgenic corn (claimed).    DETAILED DESCRIPTION - A primer set for qualitative and high-throughput multiplex detection of transgenic corn comprising specific primer pairs for transgenic corn BT176, TC1507, and MON863, is new. The specific primer sequences are as follows: BT176 comprising forward primer comprising a 52 base pair sequence (SEQ ID NO: Not defined) fully defined in the specification and reverse primer comprising a base pair sequence of: tgggcgtggtatcgactttatag (SEQ ID NO: Not defined), MON863 comprising forward primer comprising a 53 base pair sequence (SEQ ID NO: Not defined) fully defined in the specification and reverse primer comprising a base pair sequence of: actgtcggcagaggcatcttgaa (SEQ ID NO: Not defined), and TC1507 comprising a forward primer comprising a 53 base pair sequence (SEQ ID NO: Not defined) fully defined in the specification and reverse primer comprising a base pair sequence of: gccttgacttaatcgcacccatc (SEQ ID NO: Not defined). An INDEPENDENT CLAIM is included for droplet PCR coupled denaturing gradient gel electrophoresis (DGGE) analysis method for multiplex detection of transgenic corn, which involves extracting test sample DNA, adding micro-drop PCR aqueous phase to the micro-drop PCR oil phase, and mixing the materials for 1.5 minutes, where the diameter of the droplets is 1-8 mu m and aqueous droplet PCR components comprise 300-500 nM forward primer, 300-500 nM reverse primer, 10X PCR Buffer, 8-12 g/l bovine serum albumin, 0.1-0.3 mM deoxyribonucleotide triphosphate (dNTP) and 109 PCR template, performing PCR amplification of the template, where the PCR reaction include predenaturation at 94 degrees C for 5 minutes, denaturation at 94 degrees C for 30 seconds, annealing at 61-65 degrees C for 30-60 seconds and extension at 72 degrees C for 30 seconds, collecting the reaction solution, centrifuging the solution, locating the oil phase in upper layer of the centrifuge tube, the aqueous phase in the lower layer of the centrifuge tube, collecting lower aqueous phase to obtain purified microsphere PCR, subjecting the purified microtiter PCR products to DGGE electrophoresis, and visualizing the product under UV light. 
主权项：1.一种用于转基因玉米定性、高通量多重检测的引物组，该引物组包括转基因玉米BT176、TC1507和MON863的特异性引物对，具体引物序列如下： 玉米品系 引物序列(5’to3’) BT176： 正向引物： CGCCCGCCGCCGCGGCGGCGGGGCGGGGGCGTG GGTTTCTGGCAGCTGGACT； 反向引物：TGGGCGTGGTATCGACTTTATAG； MON863： 正向引物： CGCCCGCCGCCGCGGCGGCGGGGCGGGGGCTTG GTTCGGAGAGCACTTGTTGG； 反向引物：ACTGTCGGCAGAGGCATCTTGAA； TC1507： 正向引物： CGCCCGCCGCCGCGGCGGCGGGGCGGGGGCCAC CCACCGGAACAAAATGTAAC； 反向引物：GCCTTGACTTAATCGCACCCATC。
申请号：CN201711210493.1
专利号：CN107723346A
公开/公告号：CN107723346A
申请/专利权人：上海市农业科学院;
发明/设计人：唐雪明; 赵凯; 赵笑; 范小瑞; 杜亚楠; 史斌; 赵桓震; 赵斌安; 潘磊; 张琪; 张晓霞
分类号：C12Q1/686; C12Q1/6895; C12N15/11;
主分类号：C12Q1/686
法律状态：实质审查
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