中文摘要：本发明涉及分子生物学及生物检测技术领域，具体公开了基于环介导等温扩增消光技术的转基因玉米MON810检测方法。本发明针对转基因玉米MON810转化事件的3’端基因的6个区域设计4条特异性引物，其上下游外引物序列如SEQ?ID?NO.1?2所示，其上下游内引物序列如SEQ?ID?NO.3?4所示。利用环介导等温扩增消光技术使用所述引物对样品的DNA进行扩增，通过反应颜色或吸光值的变化判断是否有扩增产物，从而检测转基因玉米MON810。本发明方法具有灵敏度高、特异性强、操作简单、可肉眼判定结果等优点，为转基因玉米MON810检测提供了有效、快速的检测方法。
外文摘要：NOVELTY - Primer comprising all of fully defined 35-57 nucleobases (SEQ ID NO: 1-4) given in specification, is new.
USE - Primer, in the form of kit, used in loop-mediated isothermal amplification for detecting transgenic maize MON810 (all claimed).
ADVANTAGE - The product has high sensitivity, strong specificity, simple operation, and short detection time. The results can be easily judged by naked eye.
DETAILED DESCRIPTION - An INDEPENDENT CLAIM is included for detection of transgenic maize MON810 comprising extracting genome template of sample to be tested; mixing hemin working solution and primer comprising all of fully defined 35-57 nucleobases (SEQ ID NO: 1-4) given in specification, adding 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) coloring solution and hydrogen peroxide to obtain colored mixture, and measuring absorbance at 419 nm; performing loop-mediated isothermal amplification by mixing colored mixture and sample genome template, and reacting at 60-65 degrees C for 40-60 minutes; and determining absorbance at 419 nm, and comparing results from second step.
TF TECHNOLOGY FOCUS - BIOTECHNOLOGY - Preferred Components: The colored mixture further comprises 1.4-2.0 mM deoxynucleotide, 3-8 mM magnesium sulfate, 0.6-0.8 M betaine, 3.6-10.0 U Bst DNA polymerase large fragment, 1xThermopol buffer, and 2 mu L sample genome template. The hemin working solution comprises hemin, 10 mM monosodium phosphate, 100 mM potassium chloride, 2 mM magnesium chloride, and 0.003 wt.% Triton X-100 (RTM: polyethylene glycol tert octylphenyl ether).
主权项：基于环介导等温扩增消光技术的检测转基因玉米MON810的引物，其特征在于，所述引物包括上游外引物、下游外引物、上游内引物和下游内引物，其序列分别如SEQ?ID?NO.1?4所示。
申请号：CN201610949684.9
专利号：CN106367525-A
公开/公告号：CN106367525-A
申请/专利权人：中国农业大学
发明/设计人：黄昆仑;许文涛;徐瑗聪;王晨光;罗云波;陈冰
分类号：C12Q1/68;C12N15/11
主分类号：C12Q1/68
法律状态：实质审查
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