中文摘要：本发明提供一种基于环介导等温扩增消光技术的转基因玉米59122检测方法，首先针对转基因玉米59122转化事件5’端基因的部分区域设计一套LAMP引物组合，包括59122?F3/B3和59122?FIP/BIP(Seq?ID?No:1?4)，在所述引物组合基础上结合环介导等温扩增消光技术检测转基因玉米59122转化事件。本方法具有灵敏度高、特异性强、操作简单，结果易于观察等优点，为转基因玉米59122检测提供了有效、快速的检测方法，非常适于基层检测使用。
外文摘要：NOVELTY - Loop mediated isothermal amplification (LAMP) primer combination comprises outer forward primer 59122-F3 comprising a 36 base pair sequence (SEQ ID NO: 1) fully defined in the specification, outer reverse primer 59122-B3 comprising a 36 base pair sequence (SEQ ID NO: 2) fully defined in the specification, inner forward primer 59122-FIP comprising a 64 base pair sequence (SEQ ID NO: 3) fully defined in the specification and inner reverse primer 59122-BIP, which are designed with respect to 5' end sequence of transgenic corn 59122 transformation events.
USE - The LAMP primer combination is useful in kit for detecting transgenic corn 59122 (all claimed).
ADVANTAGE - The primer combination detects transgenic corn 59122 simply and rapidly with high sensitivity and strong specificity, and is suitable for substrate detection.
DETAILED DESCRIPTION - Loop mediated isothermal amplification (LAMP) primer combination comprises outer forward primer 59122-F3 comprising a 36 base pair sequence (SEQ ID NO: 1) fully defined in the specification, outer reverse primer 59122-B3 comprising a 36 base pair sequence (SEQ ID NO: 2) fully defined in the specification, inner forward primer 59122-FIP comprising a 64 base pair sequence (SEQ ID NO: 3) fully defined in the specification and inner reverse primer 59122-BIP comprising a 58 base pair sequence (SEQ ID NO: 4) fully defined in the specification, which are designed with respect to 5' end sequence of transgenic corn 59122 transformation events. INDEPENDENT CLAIMS are included for the following:
(1) kit comprising the LAMP primer combination; and
(2) method for detecting transgenic corn 59122 based on extinction LAMP technology, involves grinding sample into powder, weighing, extracting genomic DNA using cetyltrimethylammonium bromide method, performing chromogenic reaction, observing color change in the reaction solution through naked eyes, or using UV spectrophotometer to measure optical! density (OD) value of the reaction solution at 419 nm, amplifying the genomic DNA using the LAMP primer combination, and analyzing the amplified products.
主权项：用于检测转基因玉米59122的LAMP引物组合，其特征在于，所述引物组合根据转基因玉米59122转化事件5’端序列设计，包括：外侧正向引物59122?F3：5’?CTGGGAGGGAGGGAGGGGCACCTGTGATTGGCTCAT?3’；外侧反向引物59122?B3：5’?CTGGGAGGGAGGGAGGGGCTGAGTGGCTCCTTCAAC?3’；以及内侧正向引物59122?FIP：5’?TGTCGTTTCCCGCCTTCAGTTTCTGGGAGGGAGGGAGGGGAGGGACGGAAGAAAGAGTGAAGGG?3；’内侧反向引物59122?BIP：5’?AGGGAGTCACGTTATGACCCCCCTGGGAGGGAGGGAGGGGCGGTTCTGTCAGTTCCAA?3’。
申请号：CN201610907576.5
专利号：CN106498045-A
公开/公告号：CN106498045A
申请日：2016-10-18
公开/公告日：2017-03-15
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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