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Method for purifying mulberry phytoplasma genome

A purification method and genome technology, applied in the field of purification of mulberry phytoplasma DNA, can solve the problems of increased purification difficulty, inapplicability, and high genome content requirements

Inactive Publication Date: 2016-07-06
ANKANG UNIV
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Problems solved by technology

However, this method has high requirements for the genome content of phytoplasma, and it is not suitable for plants with high lignification. Generally, dodder is transferred to herbaceous plants, which greatly increases the difficulty of purification.
However, the method of directly purifying phytoplasma from the total genome of diseased plants by enzymatic reaction has not been reported.

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  • Method for purifying mulberry phytoplasma genome
  • Method for purifying mulberry phytoplasma genome
  • Method for purifying mulberry phytoplasma genome

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Embodiment Construction

[0024] The present invention is further described below in conjunction with the examples, but the present invention is not limited to the following examples.

[0025] A kind of purification method for mulberry phytoplasma genome, comprising the following steps:

[0026] Step 1, sample processing, because the phloem sieve tube cells of plants do not have nuclei, the genome content of mulberry is low, and the content of phytoplasma is high. Use a scalpel to take 1 g of the phloem of the branches of the diseased plant infected with phytoplasma of mulberry, put it into a clean mortar, add liquid nitrogen to freeze and grind it evenly;

[0027] Step 2: Extract genomic DNA. Compare the three genomic extraction methods of spin column, magnetic beads and ethanol precipitation. The magnetic bead method can extract the longest fragment, which can reach more than 50kbp. In order to improve the removal efficiency of the host protein in the next step, magnetic The genome of the sample was...

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Abstract

The invention discloses a method for purifying mulberry phytoplasma genome. By using the characteristic that a phytoplasma has a cell membrane and the genome of the phytoplasma is semi-autonomously copied, and adopting a DNA methylation conjugated protein, the mulberry genome is removed from purified mulberry whole genomes infected by phytoplasma, and semi-quantitative and quantitative PCRs (polymerase chain reactions) prove that the obtained DNA is relatively high in purity and is a mulberry phytoplasma genome DNA relatively high in concentration, so that the problems of low phytoplasma content and difficult pulse electrophoresis purification of woody plants are solved, the effect of the method disclosed by the invention is tested by using quantitative PCR, and the obtained phytoplasma genome is relatively high in concentration and proportion and can be used for PCR experiments and high-throughput sequencing.

Description

technical field [0001] The invention relates to a method for purifying mulberry phytoplasma genome, in particular to a method for purifying mulberry phytoplasma DNA that requires high-purity genomes such as direct sequencing. Background technique [0002] In 1967, Doi et al. first discovered that mulberry yellowing type atrophy disease, Paulownia arbusculosis and other plant arbusculosis diseases were caused by parasitism of plant mycoplasma. Mycoplasma was observed in the phloem sieve tube. In 1998, Qiu Bingsheng et al. amplified the 16SRNA fragments of Phellinus lutea atrophy and atrophy. Aster yellow group. In 1992, the mycoplasma was officially named Phytoplasma. Phytoplasma belongs to the genus Molluscum. The cells are usually round or oval, without a cell wall, with a monolayer membrane outside the cytoplasm, and there are granular ribosomes, soluble proteins, RNA and metabolites in the cytoplasm, and the genome is double. Chain closed circular DNA, currently known ...

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2521/327C12Q2563/143C12Q2523/308
Inventor 孔卫青杨金宏
Owner ANKANG UNIV
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