Leaf direct PCR (polymerase chain reaction) method and application of method to transgene plant detection

Abstract

The invention discloses a PCR (polymerase chain reaction) amplification method and application of the method. The PCR amplification method provided by the invention belongs to a method for directly adopting plant leaves as templates and adopting Phire hot starting II DNA (deoxyribonucleic acid) polymerase for PCR amplification. The leaves are tender leaves. The PCR amplification method provided by the invention is simple, the speed is high, the practicability is high, and the obtained PCR detection results are accurate and reliable. Compared with the traditional method for taking tender leaves for DNA extraction and PCR detection after the seed sprouting, the method has the advantages that the template DNA preparation step is omitted, and the detection time and the cost are greatly reduced. The method provided by the invention has great application prospects and space in the transgene plant detection and new species culture.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a PCR amplification method and its application, in particular to a method for performing PCR amplification using leaves directly as a template, and the application of the method in the detection of transgenic plants. Background technique [0002] At present, there are many methods for the detection of transgenic plants, mainly based on different detection purposes, which can be divided into the following categories: First, the purpose of detection is to identify whether the exogenous gene is integrated or not and the number of integrated copies. Multiplex PCR (Multiplex PCR, also known as MPCR), touchdown PCR (Touchdown PCR, TD-PCR), inverse PCR (inverse PCR, IPCR), and Southern hybridization detection methods, etc. Secondly, the detection and identification of whether the exogenous gene is transcribed in the transformed plant can be detected by methods such as Northern hybridization a...

AI Technical Summary

Problems solved by technology

Especially in the process of backcross breeding of transgenic plants, a large number of samples need to be tested, and how to ensure that the nucleic acids in the samples are not degraded during the long-distance transportation of a large number of leaves obtained from the field is a very difficult problem. However, it often consumes a lot of manpower, material resources and financial resources to carry out the transportation and preservation of the leaves and the detection results are not ideal.

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