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Detection kit and detection method for chicken miRNA-1606 gene polymorphism

A gene polymorphism and detection kit technology, applied in the field of molecular genetics, can solve the problems of unfavorable popularization and application, expensive equipment, high cost of gene chips, etc., achieving sensitive throughput, short detection time, and speeding up the breeding of fine varieties. Effect

Inactive Publication Date: 2015-01-21
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cost of gene chips is high, and the equipment required is expensive, which is not conducive to popularization and application

Method used

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  • Detection kit and detection method for chicken miRNA-1606 gene polymorphism
  • Detection kit and detection method for chicken miRNA-1606 gene polymorphism
  • Detection kit and detection method for chicken miRNA-1606 gene polymorphism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] In this embodiment, the chicken miRNA-1606 gene polymorphism detection kit includes primer pair P2 (PCR amplification primer) and primer P3 (single base extension primer),

[0044] Primer pair P2 is:

[0045] F Primer: 5'-ACGTTGGATGAACAAAGCAGCGTGAATGGTC-3',

[0046] R Primer: 5'-ACGTTGGATGTTCAGGACTGTTCTGTTGCC-3';

[0047] Primer P3 is: 5'-GTTGCCCCCTCTTTCAGT-3'.

Embodiment 2

[0049] The chicken miRNA-1606 gene polymorphism detection kit in this example includes 500nM primer pair P2480μL, 500nM single-base extension primer P3480μL, 25mM dNTP mix48μL, 5U / μL Hotstar Taq48μL, iPLEX single-base extension enzyme 19.68μL, 10 ×PCR buffer300μL, 25mM MgCl 2 156 μL, 888 μL ultrapure water, 96 μL iPLEX termination reaction Mix, 240 μL 1U / μL SAP enzyme, 81.6 μL 10×SAP buffer;

[0050] Primer pair P2 is:

[0051] F Primer: 5'-ACGTTGGATGAACAAAGCAGCGTGAATGGTC-3',

[0052] R Primer: 5'-ACGTTGGATGTTCAGGACTGTTCTGTTGCC-3';

[0053] Primer P3 is: 5'-GTTGCCCCCTCTTTCAGT-3'.

Embodiment 3

[0055] The detection method of chicken miRNA-1606 gene polymorphism in the present embodiment, comprises the following steps:

[0056] 1. Material preparation

[0057] Animal material: Gushi chicken-Anka chicken F2 generation resource group.

[0058] Gushi chicken is a well-known excellent local chicken breed in my country, which belongs to the type of yellow chicken. It is centered in Gushi County, and it is naturally formed after long-term closed breeding under the special ecological environment and breeding conditions. Gushi chicken has the characteristics of high egg production, large eggs and thick shells, and stable genetic performance. It has the following outstanding excellent characters: resistance to rough feeding, strong disease resistance, suitable for wild grazing and free-range breeding; second, tender meat, delicious meat, mellow soup, rich nutrition, and strong nourishing effect; third, hen production There are many eggs, the eggs are big, the egg whites are ...

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Abstract

The invention discloses a detection kit and a detection method for chicken miRNA-1606 gene polymorphism, and belongs to the technical field of molecular genetics. The detection method comprises the steps of by using chicken whole genome DNA as a template, designing a primer to amplify a target fragment including a miRNA-1606 precursor, purifying and sequencing to know that the 35th site of the chicken miRNA-1606 gene has C or A nucleotide polymorphism; then designing a primer to amplify gene segments containing polymorphism sites and extending reactions by adopting matrix assistant laser desorption ionization-time of flight mass spectrometry in combination with a single primer, and carrying out single SNP site detection on a lot of samples. The detection method has the advantages of high accuracy, sensitive throughput, short detection time and high ratio of performance to cost, can be used for assistant selection and molecular breeding of the chicken, and has important effects for increasing growth traits of the chicken.

Description

technical field [0001] The invention specifically relates to a chicken miRNA-1606 gene polymorphism detection kit and detection method, belonging to the technical field of molecular genetics. Background technique [0002] In the genetic breeding of livestock and poultry, the application of modern breeding technology of molecular markers is an effective method to speed up the breeding of fine breeds and improve the genetic quality of the population, thereby increasing the advanced and effective method of livestock and poultry production performance. The application of molecular marker breeding is first to screen and detect genetic markers closely related to economic traits of livestock and poultry at the DNA level, secondly to establish a rapid detection method for its gene polymorphism, and then to realize genetic marker-assisted selection and early diagnosis selection. As an important economic poultry, chicken is also an important model organism in scientific research. Grow...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6888C12Q2600/124C12Q2600/156C12Q2533/101
Inventor 康相涛李红孙桂荣刘小军蒋瑞瑞田亚东李国喜韩瑞丽闫峰宾王彦彬李转见
Owner HENAN AGRICULTURAL UNIVERSITY
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