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Molecular specificity labeled primers of mushroom Cr02 strain and test method

A technology of labeled primers and specificity, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc. It can solve the problems of unsatisfactory identification of mushroom strains and achieve short detection time and high accuracy Effect

Inactive Publication Date: 2010-12-15
ZHEJIANG FORESTRY ACAD
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Some PCR-based molecular marker technologies such as RAPD (Random Amplified Polymorphic DNA, Random Amplified Polymorphic DNA), AFLP (Amplified Fragment Length Polymorphism, Amplified Fragment Length Polymorphism) have been used in edible fungi in the 1990s Classification and identification of strains, but these marks are not ideal for the identification of mushroom strains

Method used

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  • Molecular specificity labeled primers of mushroom Cr02 strain and test method
  • Molecular specificity labeled primers of mushroom Cr02 strain and test method
  • Molecular specificity labeled primers of mushroom Cr02 strain and test method

Examples

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Embodiment 1

[0043] (1) Mycelium culture and genomic DNA extraction: transfer the low-temperature preserved mushroom strains to the PDA slope (PDA slope medium: take 200 grams of peeled potatoes, cut into small pieces, add 1000 ml of water, 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, 15 grams of agar, dissolve and distribute, sterilize at 121°C for 30 minutes, cool to a slope), and incubate at 25°C. After 14 days, receive 100ml PD liquid medium (PD liquid medium: take 200 grams of peeled potatoes, cut into small pieces, add 1000 ml of water and boil for 30 minutes, filter out the potato pieces, add water to the filtrate to make up to 1000 ml, add glucose 20 G, 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 the hyphae, and stored in the refrigerator at -20°C for later use. The genomic DNA was extracted using the SDS-CTAB...

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Abstract

The invention provides a molecular specificity labeled primer for shiitake mushroom Cr02 strains and a method for identifying and testing the shiitake mushroom Cr02 strains by utilizing the primer. The primer sequence is: the upstream primer: 5'-AGGCAAACTGGTTCCATGATA-3' and the downstream primer: 5'-CTGTGAAGTCA T TACAGTCGC-3'. The beneficial effect of the molecular specificity labeled primer liesin that the testing method of the invention has the advantages of short testing time and high accuracy by comparing with the conventional morphological test, the antagonistic test and the fruiting test. The testing time of the invention is required only one to three days, while the conventional antagonistic test requires at least two weeks and the fruiting test requires at least three months.

Description

(1) Technical field [0001] The invention relates to a molecular specific marker primer for the lentinus edodes Cr02 strain, and a method for identifying and detecting the lentinus edodes Cr02 strain by using the primer. (2) Background technology [0002] Lentinula edodes (Berk.) Pegler originated in the temperate-subtropical regions of Asia and Australia. Shiitake mushrooms are famous all over the world for their nutrition, medical value and unique taste, especially in China and other Southeast Asian countries. China is the first country to cultivate shiitake mushrooms, and currently the output of shiitake mushrooms has reached 70% of the world's. Lentinus edodes strains are the most important means of production in the production of lentinus edodes. At present, there are more than 100 kinds of Lentinus edodes strains used in commercial cultivation in China. In the production of lentinus edodes, some lentinus edodes strains with high-yield and high-quality cultivation character...

Claims

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Application Information

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