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Specific primers, probes, kits and methods for detecting barley components

A kit and specific technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problem of inability to distinguish barley from other species, achieve short time, accurate results, and identification The results are accurate

Active Publication Date: 2021-01-05
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the technologies of the above two patents can only be used to identify barley internal varieties or wild species and cultivated species, and cannot distinguish barley from other species

Method used

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  • Specific primers, probes, kits and methods for detecting barley components
  • Specific primers, probes, kits and methods for detecting barley components
  • Specific primers, probes, kits and methods for detecting barley components

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0061] Example 2 Probe

[0062] This embodiment provides a probe for detecting barley components. The probe designed for the ACPS gene of barley is as follows, as shown in SEQ ID NO: 3 in the sequence listing: the sequence of the probe is: 5'-TCGCGCCATTCAAGCCGTGATACGTC -3'.

Embodiment 3

[0063] Example 3 Common PCR Detection Kit

[0064] This embodiment provides a test kit for detecting barley components, each of which includes a separately packaged primer solution part and a PCR reaction solution part; the primer solution part contains the The specific primer; the PCR reaction solution part contains amplification buffer, dNTPs, Taq DNA polymerase and sterile ultrapure water for PCR amplification, and the formula of each kit is:

[0065] Upstream primer, 10μM, 0.5-1.5μL;

[0066] Downstream primer, 10μM, 0.5-1.5μL;

[0067] Taq DNA polymerase, 5U / μL, 0.2~1.2μL;

[0068] 10×Taq DNA polymerase buffer (containing 10mmol / L Mg 2+ ), 2.5μL;

[0069] dNTPs, 2.5mM, 0.25-1μL; add sterile double distilled water to a total volume of 25μL.

Embodiment 4

[0070] Embodiment 4 real-time fluorescent PCR detection kit

[0071] This embodiment provides a test kit for detecting barley components, each of which includes a separately packaged primer solution part and a PCR reaction solution part; the primer solution part contains the The specific primers and the probe described in Example 2; the PCR reaction solution part contains 2 × TaqMan Universal PCR Master Mix containing dNTPs, Taq DNA polymerase and sterile ultra- Pure water, the formula of each said kit is:

[0072] 2×TaqMan Universal PCR Master Mix 12.5μL;

[0073] Upstream primer (10 μM) 1.0 μL;

[0074] Downstream primer (10 μM) 1.0 μL;

[0075] Probe (10 μM) 0.5 μL;

[0076] Add sterile double distilled water to a total volume of 25 μL.

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Abstract

The invention provides a specific primer, a probe, a kit and a method for detecting barley component. A large number of primers are designed based on FDH, GLU2a, RBCL, ACPS and UDPg genes of barley, and primers and probes with species specificity are screened out through a large number of primer screening researches. When the primers and the probes are used for detecting barley component in a sample to be detected, barley and other species adulterants can be accurately separated, the absolute detection sensitivity reaches 0.2 copies / microliter, the relative sensitivity can reach 0.5%, and theidentification result is accurate, stable and reliable.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to a specific primer, probe, kit and method for detecting barley components. Background technique [0002] With the extensive research on genetically modified products and their wide application in daily life, the safety issues brought by genetically modified products to human health and living environment have aroused widespread concern and controversy in the world. Therefore, while actively researching genetically modified technology, various countries in the world have also conducted a lot of research on the detection and analysis methods of various genetically modified foods. However, due to the large number and variety of genetically modified foods, especially after many genetically modified foods are processed and stored under various conditions, the genetically modified ingredients are degraded in large quantities, making detection more difficu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2563/107C12Q2561/113
Inventor 邓婷婷黄文胜刘凤轩
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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