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Mushroom 507 bacterial AFLP quick-detecting method

A detection method and strain technology, applied in the detection field, can solve the problems affecting the rapid development of shiitake mushrooms, affecting the cultivation enthusiasm, and the loss of mushroom farmers, and achieve the effects of short detection time, high accuracy and good repeatability

Inactive Publication Date: 2007-09-26
上海市农业科学院食用菌研究所
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  • Abstract
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Problems solved by technology

As far as the country is concerned, the phenomenon of "same species with different names and different species with the same name" in the cultivation of shiitake mushrooms has not only brought great losses to mushroom farmers, but also affected their enthusiasm for cultivation, and also greatly affected the rapid development of shiitake mushrooms in China. and, along with the appearance of large-scale industrialized cultivation methods, the requirements for the quality of the cultivated strains of shiitake mushrooms are getting higher and higher, and it is necessary to develop a more simple, fast and accurate strain identification technology to ensure that each batch of species is used. accurate

Method used

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  • Mushroom 507 bacterial AFLP quick-detecting method

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Embodiment

[0038] Mycelia culture and extraction of genomic DNA

[0039] Adopt PDY liquid medium (20% of potato, 2% of glucose, 0.1% of yeast extract), 150rpm, 25 ℃ 15 days shake flasks to cultivate mycelia of Lentinus edodes 507 strain. The mycelium was collected by filter membrane filtration, washed twice with sterile water, dried on filter paper, labeled and stored in a -20°C refrigerator for later use. The DNA was extracted using the CTAB method, and the quality of the DNA was detected by 0.8% agarose gel electrophoresis. The concentration and relative purity of the DNA were measured with a UV spectrophotometer BECKMAN DU 640, and the DNA concentration of all samples was adjusted to 100ng / ul.

[0040] DNA double digestion

[0041] The sample DNA was digested with MseI and EcoRI. DNA sample digestion system includes (total volume 20μl):

[0042] ddH2O 14.8μl

[0043] 10×Buffer 2.0μl

[0044] 10U / μl MseI 0.25μl

[0045] 10U / μl EcoRI 0.25μl

[0046] 100×BSA 0.2μl

[0047] 100ng / ...

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Abstract

The invention discloses a rapid AFLP detecting method of mushroom 507 bacterium, which comprises the following steps: extracting bacterial culture and genome DNA; cutting DNA through Msel and EcoR1; connecting DNA segment through MseI adapter and EcoRI adapter; preaugumenting DNA enzyme-cut product; augumenting through AFLP selectively; detecting through genetic analyzer.

Description

technical field [0001] The invention belongs to the field of detection methods, in particular to a rapid detection method for mushroom 507 strain AFLP Background technique [0002] The output of shiitake mushrooms in my country has grown rapidly from 19,500 tons in 1983 to 2.42 million tons in 2005, accounting for more than 70% of the total output of shiitake mushrooms in the world. Some experts predict that due to the rapid development of China's shiitake mushroom industry, it will become the edible fungus with the highest output in the world in the past 10 years. Chinese shiitake mushrooms have attracted the attention of people in the world's mushroom industry for their astonishing development speed, high-quality quality and low cost. Chinese shiitake mushrooms have become popular all over the world. [0003] The contribution rate of high-quality strains to the yield and quality of shiitake mushrooms is very important, which determines the important position of shiitake mu...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04G01N27/447
Inventor 陈明杰谭琦尚晓冬宋春艳卓英潘迎捷
Owner 上海市农业科学院食用菌研究所