A method for constructing DNA fingerprints of hop varieties using ssr molecular marker technology

A technology of DNA fingerprinting and molecular markers, which is applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve problems such as questionable authenticity, and achieve low-cost effects

Active Publication Date: 2020-05-29
TSINGTAO BREWERY
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Problems solved by technology

However, there are many reports on the identification of hop varieties at home and abroad, but the authenticity remains to be discussed. Therefore, how to use SSR technology to identify the authenticity of hop varieties at home and abroad, and ensure that the quality of purchased hops meets the needs of brewing from the source, which will It is an important topic that should be studied by those skilled in the art

Method used

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  • A method for constructing DNA fingerprints of hop varieties using ssr molecular marker technology
  • A method for constructing DNA fingerprints of hop varieties using ssr molecular marker technology
  • A method for constructing DNA fingerprints of hop varieties using ssr molecular marker technology

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Embodiment 1

[0031] 1. Materials and instruments

[0032] Materials: 8 hop varieties, of which 5 are domestic hops and 3 are imported hops. See Table 1 for the variety names and origins.

[0033] Table 1 Eight hop varieties

[0034]

[0035] Take the fresh hop cones of the above hop varieties, place them in an oven at 50°C and dry them to a moisture content of 6%, to obtain corresponding hop samples, and store them at 4°C for later use.

[0036] Reagents: Plant DNA Extraction Kit, Qiagen, Germany; Taq DNA polymerase, dNTPs, ThermoFisher, USA; 30% (w / v) acrylamide solution, 5×TBE buffer, ammonium persulfate, tetramethylethylenediamine , DNAMarker, Sangon Bioengineering (Shanghai) Co., Ltd.; primers were synthesized by Shanghai Yingjun Biotechnology Co., Ltd.

[0037] Instruments: PCR instrument, PAC3000 electrophoresis instrument, American Bio-Rad Company; centrifuge, German Sigma Company; high-throughput tissue grinder, Beijing Tomogen Company; UV spectrophotometer, American GE Compan...

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Abstract

The invention provides a method for constructing DNA fingerprints of hop varieties by using a simple sequence repeat (SSR) molecular marker technology, belonging to the technical field of biology. Themethod can be used for carrying out authenticity identification on eight main hop varieties. The method is characterized by comprising the steps of extracting deoxyribonucleic acid (DNA) of test samples, selecting 4 pairs of SSR primers to perform polymerase chain reaction (PCR) amplification on the different hop varieties, carrying out non-denaturing polyacrylamide gel electrophoresis, performing silver staining, and carrying out data analysis and map construction. The method can be used for constructing the fingerprints of the main hop varieties used by beer enterprises in China, thus separately giving a fingerprint code to all the varieties at the molecular level for accurately indicating the identities of the varieties; the authenticity identification of the eight hop varieties can berealized by means of the standard fingerprints.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing DNA fingerprints of hop varieties by using SSR molecular marker technology. Background technique [0002] Beer is a low-alcohol wine full of carbon dioxide brewed from malt, hops, and water through yeast fermentation. Hops are an essential raw material for beer brewing, and different hop varieties will give beer different flavor characteristics, so choosing the right hops in the beer brewing process is very important for beer brewing. [0003] Currently, hop variety identification techniques include traditional morphological identification, chemical composition analysis and DNA molecular markers. The traditional hop variety identification method is based on the morphological and anatomical characteristics of hop cultivars to identify hop varieties, but it is unable to distinguish hop particle samples after deep processing. However, based on che...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6858
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 尹花董建军张志军岳杰杨梅胡淑敏周月南刘明丽张磊张翠
Owner TSINGTAO BREWERY
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