Detection of CaMV 35S promoter and nos terminator by adopting RPA (Recombinase Ploymerase Amplification) technology

Abstract

The invention provides an RPA sifting and detecting method for transgenic crops containing P-35S and T-nos elements, and particularly discloses a primer and probe combination applicable to the identification of transgenic plants containing transgenic elements, namely a CaMV 35S promoter and a nos terminator by adopting an RPA technology. The forward primer sequences of the primer and probe combination are shown in SEQ ID No.1 and SEQ ID No.4, the reverse primer sequences of the primer and probe combination are shown in SEQ ID No.2 and SEQ ID No.5, and the probe sequences of the primer and probe combination are shown in SEQ ID No.3 and SEQ ID No.6. In addition, the invention further discloses a method for identifying transgenic crops containing the two kinds of elements, and the method comprises the following steps: extracting the DNA of a sample to be detected as a template, and using the primer to perform RPA rapid amplification and real-time fluorescence detection, if an obvious amplification curve is obtained, the fact that the DNA of the detected sample contains P-35S and T-nos transgenic elements is proved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to rapid identification of CaMV 35S promoter (P-35S) and / or nos terminator (T- nos) components, specifically related to the combination of primers and probes and their monitoring methods. Background technique [0002] At present, DNA amplification is the main method of nucleic acid detection. The commonly used PCR detection requires sophisticated instruments and cumbersome test procedures, which is difficult to meet the requirements of on-site detection in non-laboratory environments. In recent years, nucleic acid constant temperature amplification technology has been greatly developed. Compared with traditional PCR, nucleic acid constant temperature amplification technology does not require expensive PCR equipment, and can quickly amplify the target fragment in a short time. It is simple, fast, sensitive, etc. advantage. RPA technology is developed by simulating DNA replication in orga...

AI Technical Summary

Problems solved by technology

In the reported detection methods of genetically modified plants, the PCR instrument is mainly used for routine detection in the laboratory, and this method cannot further meet the rapid detection of genetically modified products.

At present, RPA technology has not been used to screen and identify transgenic plants at home and abroad

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