Specific primer, probe, test kit and method for detecting sugarcane components

A specificity and kit technology, applied in kits, specific primers and probes for detecting sugarcane components, can solve the problem of inability to distinguish sugarcane from other species, and achieve small molecular weight, high amplification efficiency and high sensitivity Effect

Active Publication Date: 2020-06-26
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the technology of the above-mentioned patent can only be used to identify the genetic relationship of different samples within the species of sugarcane, and cannot distinguish sugarcane from other species

Method used

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  • Specific primer, probe, test kit and method for detecting sugarcane components
  • Specific primer, probe, test kit and method for detecting sugarcane components
  • Specific primer, probe, test kit and method for detecting sugarcane components

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0066] Example 2 Probe

[0067] This embodiment provides a probe for detecting sugarcane components. The probe designed for the ACPS gene of sugarcane is as follows, see SEQ ID NO: 3 in the sequence listing, the sequence of the probe is: 5'-CGATCGATGTTCTGGGTGCTGAGTGCC -3'.

Embodiment 3

[0068] Example 3 Common PCR Detection Kit

[0069] The present embodiment provides a kind of test kit for detecting sugarcane components, and each said kit includes a primer solution part and a PCR reaction solution part of independent packaging; 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:

[0070] Upstream primer (10 μM) 1.0 μL;

[0071] Downstream primer (10 μM) 1.0 μL;

[0072] Taq DNA polymerase, 5U / μL, 0.4μL;

[0073] dNTPs, 10mM, 0.5μL;

[0074] 10×Taq DNA polymerase buffer (containing 10mmol / L Mg 2+ ), 2.5 μL;

[0075] Make up to 25 μL with sterile ultrapure water;

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

Embodiment 4

[0077] Embodiment 4 real-time fluorescent PCR detection kit

[0078] The present embodiment provides a kind of test kit for detecting sugarcane components, and each said kit includes a primer solution part and a PCR reaction solution part of independent packaging; The specific primers and the probe described in Example 2; the PCR reaction solution part contains 2×TaqManUniversal PCR Master Mix of dNTPs, Taq DNA polymerase and sterile ultrapure water, the formula of each kit for:

[0079] 2×TaqMan Universal PCR Master Mix 12.5μL;

[0080] Upstream primer (10 μM) 1.0 μL;

[0081] Downstream primer (10 μM) 1.0 μL;

[0082] Probe (10 μM) 0.5 μL;

[0083] 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 test kit and a method for detecting sugarcane components. According to the invention, a large number of primers are designed on the basis of the ITS gene, rbcl gene, nsLTP gene and NDR1/HIN1 gene of sugarcane; meanwhile, through screening research on the large number of primers, a primer and a probe with species specificity are screened out; andthe above-mentioned primer and probe can be used for detecting the sugarcane components in a to-be-detected sample, can accurately separate sugarcane from species, can reach an absolute detection sensitivity of 0.4 copies/[mu]L and a relative detection sensitivity of 0.01%, and have accurate, stable and reliable identification results.

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 sugarcane 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 diff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/166C12Q2563/107C12Q2545/113C12Q2561/113
Inventor 邓婷婷黄文胜刘凤轩
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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