Lysis solution used for plant parasitic nematode genome DNA micro-extraction and application thereof

A lysate, genome technology

Inactive Publication Date: 2014-08-27
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome defects such as low efficiency, low success rate, complicated steps and the like in the existing plant parasitic nematode genome DNA micro-extraction technology, and provide a lysate for micro-extraction of plant parasitic nematode genome DNA and Its application, described lysate is as follows:

Method used

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  • Lysis solution used for plant parasitic nematode genome DNA micro-extraction and application thereof
  • Lysis solution used for plant parasitic nematode genome DNA micro-extraction and application thereof
  • Lysis solution used for plant parasitic nematode genome DNA micro-extraction and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, different lysates (lysate 1~4) extract the effect comparison of 5 kinds of plant parasitic nematode genomic DNAs Nematode DNA extraction:

[0030] Clean the nematodes with double distilled water, pick a single nematode and put it into a 200 μL PCR tube containing 10 μL of lysate, and then heat at 56 °C (heat for 56 °C for 1 h for rice stem nematode, 56 °C for 2 h for pine xylophilus and pseudo-pine wood nematode , Brachybody nematodes heated at 56°C for 3h), heated at 95°C for 10min, and the extracted supernatant obtained after instantaneous centrifugation can be used for molecular experiments. Set up 4 repetitions.

[0031] Evaluation of the effect of different lysates on nematode DNA extraction:

[0032] The extracted nematode DNA samples were subjected to PCR amplification of the ribosomal ITS region and detected by electrophoresis to evaluate the effect of different lysates on nematode genomic DNA extraction.

[0033] 1. Formulas of different lysates...

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Abstract

The invention provides a lysis solution for plant parasitic nematode genome DNA micro-extraction and application of the lysis solution. The lysis solution is characterized by being allowed to be directly used for plant parasitic nematode genome DNA micro-extraction without cutting, extruding and breaking, grinding and freezing and thawing nematodes, operation steps are simpler and more convenient to implement, and sample pollution and loss are reduced to the maximum degree in the DNA extraction process, extraction efficiency is obviously improved, the success rate is obviously increased, and the problems that in the prior art, efficiency is low, stability is poor, operation is difficult and the steps are numerous in the plant parasitic nematode DNA extraction process are solved effectively. Nematode DNA obtained through extraction by the adoption of the method meets the requirements of plant parasitic nematode DNA barcode identification, RFLP enzyme digestion identification, specific primer PCR identification and other molecular detection technologies, and the lysis solution can be applied and popularized in quarantine identification work of China ports, agricultural departments and forestry departments.

Description

technical field [0001] The invention relates to the field of plant parasitic nematode quarantine and identification, and provides a lysate for micro-extraction of plant parasitic nematode genome DNA and its application. Specifically, the method greatly simplifies The genome DNA extraction program of plant parasitic nematodes has significantly improved the efficiency and success rate of DNA extraction, and effectively solved the problems of low efficiency, poor stability, difficult operation, and many steps in the previous micro-extraction process of plant parasitic nematode DNA. It is suitable for ports And agriculture and forestry related quarantine and identification laboratory applications. Background technique [0002] The traditional identification methods of plant nematodes mainly rely on morphological characteristics and morphological measurements, requiring the examiners to have extremely rich experience and skills. The DNA-based molecular identification method has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68
Inventor 王金成顾建锋林宇张裕君冯洁陈先锋黄国明
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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