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Molecule-specific labeled primer and method for identifying European boletus related species

A technology for labeling primers and boletus, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of subjective judgment bias and color difference, and achieve the effect of simple methods

Inactive Publication Date: 2013-04-17
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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Problems solved by technology

However, because the color difference is easily affected by habitat, climate, soil, fruiting body growth period, physiological status, etc., it often leads to deviations in subjective judgment. For long-term preserved dry specimens, especially for re-identification of ancient specimens and type specimens Research, it is difficult to accurately identify only by color difference

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  • Molecule-specific labeled primer and method for identifying European boletus related species
  • Molecule-specific labeled primer and method for identifying European boletus related species
  • Molecule-specific labeled primer and method for identifying European boletus related species

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Embodiment 1

[0037] (1) Extraction of genomic DNA from boletus specimens: Take 0.2 g of the fruiting bodies of boletus specimens to be tested, add liquid nitrogen and grind them thoroughly, and use SDS-CTAB method to extract genomic DNA. Genomic DNA crude extract of the specimen. The crude DNA was purified by Magabio Nucleic Acid Purification Kit (Bioer, Hangzhou, China), and detected by 1.5% agarose gel electrophoresis and DNA / RNA UV spectrophotometer (GeneQuant Pro, GE Healthcare). Integrity, Purity and Concentration. OD 260 / OD 280 DNA samples >1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -20°C refrigerator for later use.

[0038] (2) Design specific PCR amplification primers. The sequence of the primer pair is upstream primer 5′-TAGCTCACAGCCACTCAATGT-3′ and downstream primer 5′-GGGATGGGACAGTCGGATAATGTT-3′, which were synthesized by Shanghai Bioengineering Technology Co., Ltd.

[0039] (3) PCR amplification with ITS-specific primers:

[0040] PCR r...

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Abstract

The invention provides a molecule-specific labeled primer for identifying European boletus related species Boletus appendiculatus and B. pseudoregius, and a method for quickly identifying the European boletus related species B. appendiculatus and B. pseudoregius by using the molecule-specific labeled primer. The molecule-specific labeled primer has sequences as follows: an upstream primer: 5'-TAGCTCACAGCCCACTCAATGT-3', and a downstream primer: 5'-GGGATGGGACAGTCGGATAATGTT-3'. The molecule-specific labeled primer can perform quick molecular Identification on the related species B. appendiculatus and B. pseudoregius; and the method is simple, quick and accurate and is an effective auxiliary method for identifying the two kinds of boletus based on morphological characteristics.

Description

(1) Technical field [0001] The invention relates to a molecular specific marker primer for distinguishing Boletus appendiculatus and B. pseudoregius, and a method for quickly distinguishing B. appendiculatus and B. pseudoregius using the primer Methods. (2) Background technology [0002] The Appendiculati Konrad & Maubl. ex Lannoy & Estadès group belonging to the Boletus genus under Boletales includes Boletus appendiculatus Schaeff. : Fr., B. subappendiculatus Dermek, Lazebn. & J. Veselsky, B. fechtneri Velen., B. pseudoregius (Huber) Estadès and B. regius Krombh. Five Boletus relatives. These five closely related species all originated in Europe, and their common characteristics are that the stipe of the fruiting body is more or less reticulated, the whole stipe is uniformly yellow, and the tubes and pores often turn blue; the flesh turns blue or Not blue, but always mild in taste; many species have a pinkish tinge at the base of the stipe. [0003] The typical feature f...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12R1/645
Inventor 丁红梅葛尔宁盛振华
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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