PCR method identified by thelephora ganbajun Zang and specific primer thereof

A dry and specific technology, applied in the direction of DNA / RNA fragments, biochemical equipment and methods, microbial measurement / inspection, etc., can solve the problems of insufficient data and high market prices, and achieve good repeatability and strong specificity Effect

Active Publication Date: 2011-05-04
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Ganba is a symbiotic fungus, which cannot be artificially cultivated at present. All commercially available commodities come from wild resources, and due to the high market price, it has led to the predatory exploitation of wild resources. Wild resources are facing great pressure. The protection and rational utilization of dry mushroom resources have become an urgent problem
Compared with other wild edible fungi with high economic value, domestic and foreign researches on Dried Mushrooms are still very few. At present, there are not many existing materials, and most of them focus on classification, strain isolation, nutrient composition determination and ecology. There is no report on the PCR method and its specific primers for bacterial identification

Method used

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  • PCR method identified by thelephora ganbajun Zang and specific primer thereof
  • PCR method identified by thelephora ganbajun Zang and specific primer thereof
  • PCR method identified by thelephora ganbajun Zang and specific primer thereof

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Experimental program
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Effect test

Embodiment 1

[0017] See Table 1 for the collection of Thelephora ganbajun Zang fruiting bodies, pure cultures of Thelephora ganbajun Zang, related species, and associated bacteria.

[0018] Table 1 Fruiting bodies, pure cultures, relative species and associated bacteria of D.

[0019]

Embodiment 2

[0021] Specific primer design.

[0022] Search GenBank (http: / / www.ncbi.nlm.nih.gov / genbank / ) for the transcriptional spacer (ITS) sequence of the genus Thelephora, use ClustalX1.8 for sequence alignment, and design according to the variable region The specific primers for Thelephora ganbajun Zang are as follows: Tg1: 5'-ACCTGTGCACCCTCTGTAGTTCC-3'; Tg2: 5'-GTCCAAGCTCATCATGGCA-3'

Embodiment 3

[0024] DNA extraction

[0025] Get about 0.1 g of the fungal samples listed in Table 1 (grown mycelium or fruiting body), transfer to a 1.5 ml centrifuge tube, add 0.30 g of white quartz sand (Sigma) and 60 ° C 2 × CTAB buffer [2 %CTAB (w / v); 100mM Tris-HCl; 1.4M NaCl; 20mM EDTA, pH 8.0] 1ml, grind thoroughly with a plastic mallet; place in a water bath at 60°C for 1 hour, then repeat with phenol:chloroform (1:1) Extract and centrifuge until a clear liquid extract is obtained; use 4°C 100% ethanol to precipitate DNA, wash the DNA pellet with 70% cold ethanol (4°C), dry the DNA pellet with a vacuum centrifugation system, and resuspend the DNA pellet in 100 μl TE buffer solution; take 5 μl DNA solution and electrophoresis on 1.5% agarose gel to detect the purity and integrity of genomic DNA; finally, store the obtained DNA sample at 4°C.

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Abstract

The invention relates to a PCR method identified by thelephora ganbajun Zang and a specific primer thereof. The Thelephora ganbajun Zang can be efficiently distinguished from other telephora and thelephora ganbajun Zang companion fungus with similar shape through DNA extraction, specificity PCR reaction and agarose gel electrophoresis detection, and a quick and accurate identification method is provided for the separation and purification of thelephora ganbajun. The invention has the advantages of high efficiency, speediness, strong specificity, good repeatability, and the like.

Description

technical field [0001] The invention relates to a PCR method for identifying Thelephora ganbajun Zang and specific primers thereof, and belongs to the technical field of using molecular biology methods to detect edible fungi. Background technique [0002] Wild edible fungi are an important forest resource and play an important role in green food development, biopharmaceuticals, environmental protection and other industries, especially some wild edible fungi distributed in forest areas, such as Tricholoma matsutake, orange Amanita caesarea, Russula virescens, Boletus edulis, Thelephora ganbajun Zang, etc. have high edible and medicinal value. In recent years, the market price has been rising. In terms of the classification of these wild edible fungi and the identification of their mycelium isolates, people mainly determine the species according to the morphology, anatomy and other characteristics of the fruiting bodies of wild edible fungi. When wild edible fungi do not prod...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 沙涛李文鹏张亚平
Owner YUNNAN UNIV
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