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Phytoplasma probe, gene chip and method for detecting phytoplasma

A gene chip and phytoplasma technology, which is applied in the high-pass detection gene chip, detects whether plant samples contain phytoplasma, and the field of phytoplasma probe, which can solve the problem of not being able to detect multiple phytoplasma with high throughput, and achieve the detection results. Output of normalized, highly specific, high-throughput assays

Inactive Publication Date: 2010-08-18
SHENZHEN POLYTECHNIC
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Problems solved by technology

[0009] The technical problem to be solved by the present invention is to provide a variety of phytoplasma probes, a gene chip containing the probes, and a method for detecting phytoplasma using the gene chip, so as to solve the problem that the prior art cannot simultaneously, rapidly and high-throughput detect multiple phytoplasma problem

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  • Phytoplasma probe, gene chip and method for detecting phytoplasma
  • Phytoplasma probe, gene chip and method for detecting phytoplasma
  • Phytoplasma probe, gene chip and method for detecting phytoplasma

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example 1

[0105] Taking the detection of Guangzhou mulberry phytoplasma as an example to further illustrate the gene chip detection sample step of the present invention, the sample detection step is as follows:

[0106] 1) Plant RNA extraction: Take 1-2g of diseased leaves, cut them into small pieces, grind them into powder with liquid nitrogen, transfer them into a sterilized 1.5mL centrifuge tube, and extract the total RNA of the samples by conventional CTAB method.

[0107] 2) Amplification of phytoplasma-specific fragments: the extracted sample DNA is labeled by PCR amplification method, and the external reference positive control plasmid samples are mixed in equal amounts and then PCR-amplified and labeled with universal primers:

[0108] (1) PCR amplification:

[0109] Specific amplification reaction system (20μL):

[0110] DNA sample 0.1μg

[0111] Amplification primer pair (5μM) 1μL

[0112] 10×PCR Buffer 2μL

[0113] 2.5mM d(AGT)TP mix 2μL

[0114] 0.25mM dCTP 2μL

[0115...

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Abstract

The invention relates to a phytoplasma probe, and comprises a gene chip of the probe and a method for detecting phytoplasma with the gene chip. The phytoplasma probe is a specific probe for 11 types of phytoplasma, which is point sampled and manufactured into the gene chip, and the method for detecting the phytoplasma comprises the following steps that: a plant DNA sample is extracted; a sample specific fragment is amplified through a universal primer and a product is marked; hybridization; the sample to be detected and to be marked is added into hybrid buffer fluid, and mixed uniformly into hybrid mixture; the mixture is dripped into a hybrid area of the gene chip, and put into a hybrid box for hybridization; and the hybridized chip is taken out and dried, scanning and data analysis are carried out, and whether phytoplasma exists in the sample is judged. The gene chip manufactured from the probe of the invention can quickly detect 11 types of phytoplasma with high efficiency and highflux.

Description

technical field [0001] The invention relates to the field of phytoplasma detection, in particular to a phytoplasma probe, a high-pass detection gene chip containing the probe and a method for detecting whether a plant sample contains phytoplasma by using the gene chip. Background technique [0002] In 1967, Japanese scientists used an electron microscope to observe for the first time that there were microorganisms in the phloem sieve tube cells of 4 kinds of plants with arbuscular symptoms, such as mulberry, petunia, potato and Paulownia, which were similar in shape and structure to animal mycoplasma. The medium is artificially cultivated and cannot infect animals, and it is called Mycoplasma-like organism (MLO). At the Ninth International Organization for Mycoplasmology (IOM) Congress in 1992, the use of phytoplasma as the new name and tentative genus name (Phytoplasmasp) for this type of prokaryote was proposed for the first time. It is a kind of prokaryotic organisms wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C40B40/06
Inventor 张丽君罗焕亮吴晓明
Owner SHENZHEN POLYTECHNIC
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