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Molecule specificity making primer of mushroom Cr02 bacterial and detection method

A labeled primer and specific technology, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of mushroom farmers’ loss, affecting cultivation enthusiasm, and affecting the rapid development of shiitake mushrooms, etc., to achieve detection Short time, high accuracy results

Inactive Publication Date: 2010-12-15
ZHEJIANG FORESTRY ACAD
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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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  • Molecule specificity making primer of mushroom Cr02 bacterial and detection method
  • Molecule specificity making primer of mushroom Cr02 bacterial and detection method
  • Molecule specificity making primer of mushroom Cr02 bacterial and detection method

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Embodiment 1

[0043] (1) Mycelia culture and extraction of genomic DNA: Transfer the mushroom strains preserved at low temperature to PDA slant (PDA slant medium: get 200 grams of peeled potatoes, cut into small pieces, add 1000 ml of water and boil for 30 minutes, filter Remove the potato pieces, add water to the filtrate to make up to 1000 ml, add 20 grams of glucose and 15 grams of agar, dissolve and subpackage, sterilize at 121°C for 30 minutes, cool to a slant), and incubate at 25°C. After 14 days, receive 100ml of PD liquid medium (PD liquid medium: take 200 grams of peeled potatoes, cut into small pieces, add 1000ml of water and boil for 30 minutes, filter out the potato pieces, add water to the filtrate to 1000ml, add glucose 20 gram, sterilized at 121°C for 30 minutes, cooled to obtain PD liquid medium), cultured with shaking at 100r / min at 25°C for 14 days, collected mycelia, and stored in a -20°C refrigerator for later use. Genomic DNA was extracted using the SDS-CTAB method, and...

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Abstract

The invention provides a molecule particularity mark primer of mushroom Cr02 strain and a method for to distinguishing and detecting the mushroom Cr02 strain by using the primer. The sequence of the primer is: an upstream primer is 5'-TCTCGATGCAAGAAACTTGAA-3', a downstream primer is 5'-TCGGTCAAAATTAGTCAGGT-3'. The invention has the main advantages that compared with conventional morphological detection, antagonism and fruiting experiment, the method has short detection time and high accuracy; the method only needs 1-3 days for detection, while the conventional antagonism needs at least two weeds for detection, and the fruiting experiment needs at least 3 months for detection.

Description

(1) Technical field [0001] The invention relates to a molecular-specific marker primer of the mushroom Cr02 strain, and a method for identifying and detecting the mushroom Cr02 strain by using the primer. (2) Background technology [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, excellent quality and low cost, and Chinese shiitake mushrooms have become popular all over the world. The contribution rate of excellent strains to the yield and quality of shiitake mushrooms is very important, which...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 吴学谦李海波魏海龙付立忠吴庆其贺亮程俊文
Owner ZHEJIANG FORESTRY ACAD
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