Micro-array chip detection method for 10 on-melon diseases and used chip probe

Abstract

The invention discloses a micro-array chip for 10 on-melon diseases, and also discloses a micro-array chip detection method for the 10 on-melon diseases by using the chip. The following steps are executed in sequence for each to-be-tested sample: 1) extracting the DNA of the to-be-tested sample; 2) performing a PCR reaction on a primer fluorescent marker; 3) mixing volume-equal PCR amplification products corresponding to at least two germs obtained by step 2); 4) chip-hybridizing and washing: denaturating one PCR amplification product corresponding to each germ obtained by step 2) or a mixture obtained by step 3) at 95 DEG C for 5min, and then ice-bathing for 5min to obtain after-pretreatment PCR amplification products; then chip-hybridizing and washing the after-pretreatment PCR amplification products, wherein the chip-hybridizing is performed at 45 DEG C for 1h; 5) scanning the chip to acquire a detection result.

Description

technical field [0001] The invention relates to the field of plant disease detection, in particular to a microarray chip detection method for 10 kinds of diseases on melons and the chip probes used. Background technique [0002] With the development of molecular biology technology, the detection technology of plant diseases has developed from traditional morphological identification to the current level of molecular detection, and has made great progress in various aspects such as detection speed, sensitivity and accuracy. The application of serological detection, common PCR detection technology, and fluorescent PCR detection technology has made the detection of diseases more advanced in terms of sensitivity, specificity, and accuracy. Requirements, the emergence of gene chip technology, the sample detection and analysis process to achieve continuous, integrated and miniaturized, and has a high-throughput, high-sensitivity detection level, can simultaneously detect a variety...

AI Technical Summary

Problems solved by technology

In Xinjiang Bureau’s melon seed export trade, there are hundreds of batches of testing requirements for these 10 diseases every year. When faced with a large number of testing samples, especially when the same sample is required to detect multiple diseases, it is difficult to achieve high throughput. Rapid simultaneous multi-disease detection, especially for fungal and bacterial diseases with a very long separation cycle and high experience requirements. It is difficult to determine positive samples and the sensitivity is not high, which greatly affects the final detection results.

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