Primer, kit and method for detecting transgenic maize NOS terminator

Abstract

The invention discloses a primer, a kit and a method for detecting a transgenic maize NOS terminator. The sequence of the Zein F of the primer is TTATGCTCCTTGGTCTTT. The sequence of the Zein R of the primer is GTAATAGGGCTGATGATTG. The sequence of the NOS F of the primer is TAATTGCGGGACTCTAAT. The sequence of the NOS R of the primer is AATCCTGTTGCCGGTCTT. The kit comprises 10 microliters of HRM reaction premix liquid, 0.2-3.4 microliters of the primer and 1 microliter of DNA (deoxyribonucleic acid) template, and water is added to reach 20 microliters. The detecting method includes: extracting a sample DNA; using the kit to perform PCR (polymerase chain reaction) amplification on the sample DNA; performing high-resolution melting curve analysis on the products after the amplification, and combining an amplification curve to judge results. The primer, the kit and the method are used for detecting maize Zein genes, namely endogenuous reference genes and a maize transcription terminator NOS, the kit is reasonable in component and proportion, convenient to use, and fast and accurate in detection, and the method is simple and fast to operate, low in cost and accurate in detecting result.

Description

technical field [0001] The present invention relates to a primer for detecting the terminator of transgenic corn NOS and endogenous gene Zein; the present invention also relates to a dual fluorescent PC detection kit including the above primers, and the present invention also relates to a detection kit for transgenic corn NOS Terminator and endogenous gene Zein method. Background technique [0002] Maize is one of the most important food crops in the world and occupies an important position in the world's agricultural production. In 2005, a major breakthrough was made in the development and promotion of genetically modified crops. For the first time, the European Commission allowed the commercial planting of genetically modified insect-resistant corn MON810 in 25 EU countries. Czechoslovakia joined the ranks of genetically modified corn planting, bringing the number of EU countries planting genetically modified crops to 5 . [0003] The development of the industrialization...

AI Technical Summary

Problems solved by technology

[0007] However, the conventional PCR method needs electrophoresis after PCR amplification, which is not only cumbersome to operate, but also easily causes pollution, and also needs to use fluorescent fuels that may cause cancer. Although the probe-based fluorescent PCR method is sensitive and accurate, the linear structure of the TaqMan probe leads to High background fluorescence, if the distance between the fluorophore and the quencher group is too close, the possibility of the probe degrading between them during the PCR amplification process will be greatly reduced, so that it will not function as a probe

On the contrary, if the fluorescent group and the quencher group are placed at both ends of the probe, the fluorescent background signal will be strengthened, which will affect the sensitivity of its detection.

TaqMan's high specificity will lead to a base that also limits its detection target to be too narrow. In addition, its synthesis cost is high, and the reagent has a short validity period and is easy to degrade, so it cannot be widely used.

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