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Primer pair and method for identifying hericium erinaceus houjie No.2

A technology of Hericium erinaceus and primer pairs is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., which can solve the problems of complex PCR amplification patterns, unsuitable for variety identification, and low specificity, etc., and achieve identification operations. Convenience, short detection time, and high specificity

Active Publication Date: 2016-05-11
HANGZHOU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, these molecular marker technologies use universal primers, and their PCR amplification patterns are not only complicated, but also poor in repeatability and specificity, so they are not suitable for variety identification. DNA fingerprinting can really be used for the accurate and rapid identification of the Hericium erinaceus species

Method used

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  • Primer pair and method for identifying hericium erinaceus houjie No.2
  • Primer pair and method for identifying hericium erinaceus houjie No.2
  • Primer pair and method for identifying hericium erinaceus houjie No.2

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Embodiment 1

[0031] An exploratory study was carried out on the molecular identification of 15 main species of Hericium erinaceus using SRAP molecular marker technology. Extract the hyphae genome DNA of each Hericium erinaceus species as a template, carry out PCR amplification with a large number of SRAP random primer pairs, and detect the amplified products by agarose gel electrophoresis, and finally screen to obtain differential bands (only Hericium erinaceus L. No. 2 has a pair of primers but the other varieties do not). The differential band was recovered by gel cutting, then connected to the cloning vector and sent for sequencing. The size of the obtained DNA fragment was 597bp, and its base sequence was shown in SEQIDNo.1. This DNA fragment was the specific DNA of Houjie 2 fragment.

[0032] A primer pair was designed for the specific DNA fragment of Houjie 2 obtained, and the sequence of the primer pair is:

[0033] Upstream primer: 5′-CAAACCGGTGTCAGGGAACTGC-3′

[0034] Downstrea...

Embodiment 2

[0038] (1) Extraction of Hericium erinaceus Genomic DNA

[0039] Take 0.03g of Hericium erinaceus mycelium to be tested, add liquid nitrogen to grind thoroughly, use a new rapid plant genome DNA extraction kit (DP3111, BioTeke, Beijing BioTeke Biotechnology Co., Ltd.) to extract genomic DNA, and obtain improved varieties of Hericium erinaceus Genomic DNA crude extract. The integrity, purity and concentration of the obtained crude genomic DNA were detected by agarose gel electrophoresis and a DNA / RNA UV spectrophotometer (Nanodrop Technologies, USA). DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -20°C refrigerator for later use.

[0040] (2) PCR amplification

[0041] Composition of PCR reaction solution: 2×PowerTaqPCRMasterMix 10μL, 10μM upstream and downstream specific primer pairs 1.5μL each, 20ng / μL template DNA 3μL, ddH 2 04 μL.

[0042] The amplification reaction was carried out on a LifeECO amplification ins...

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Abstract

The invention discloses a primer pair and method for identifying hericium erinaceus houjie No.2. The primer pair comprises an upstream primer and a downstream primer, wherein the upstream primer is 5'-CAAACCGGTGTCAGGGAACTGC-3', and the downstream primer is 5'-ACGAATTTGAATCCTCCACGTCGAA-3'. The primer pair is obtained through hericium erinaceus houjie No.2 specific molecular marker design. The primer pair has the advantages that detection time is short, accuracy is high, judgment is easy and repeatability is high when used for identifying hericium erinaceus houjie No.2 with the PCR method, and early-stage identification can be achieved quickly.

Description

technical field [0001] The invention relates to the technical field of molecular identification, in particular to a pair of primers and a method for identifying Hericium erinaceus Houjie 2. Background technique [0002] Hericium erinaceus (Hericium erinaceus), also known as Hericium erinaceus, got its name only because of its resemblance to Hericium erinaceus. Monkey mushroom, Hericium erinaceus, Hericium erinaceus, is a famous and precious traditional Chinese dish, the meat is tender, fragrant, delicious. Hericium erinaceus is a rare fungus with both edible and medicinal properties, and its fruiting body and fermented hyphae can be used as medicine. With the improvement of people's living standards, Hericium erinaceus has also entered ordinary families, especially after the "SARS", the nutritional and medicinal value of Hericium erinaceus has become more recognized, the market consumption of Hericium erinaceus has increased sharply, and the broad market demand has made Ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895C12Q2600/156
Inventor 周祖法陆娜宋吉玲
Owner HANGZHOU ACAD OF AGRI SCI
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