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Single nucleotide polymorphism, detection method and application of chicken gene

A single nucleotide polymorphism and gene technology, applied in the field of molecular genetics, can solve the problems of long time consumption, many influencing factors, and high cost of direct sequencing technology, and achieve the effect of low cost and accurate detection

Inactive Publication Date: 2015-06-17
HENAN AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, SSCP operation is cumbersome, time-consuming, and there are many influencing factors, and there are false negative problems in the experimental process, so it is not an ideal SNP detection method; the cost of direct sequencing technology is relatively high

Method used

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  • Single nucleotide polymorphism, detection method and application of chicken gene
  • Single nucleotide polymorphism, detection method and application of chicken gene
  • Single nucleotide polymorphism, detection method and application of chicken gene

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1, Extraction and Detection of Chicken Genomic DNA

[0036] 1. Extraction of Chicken Genomic DNA

[0037] Take Gushi chicken-Anka chicken resource group F 2 A total of 772 chickens were used as materials, and 5 mL of jugular venous blood was extracted from each chicken, put into a centrifuge tube, placed at an angle at room temperature for 30 minutes, and put into a centrifuge after precipitation of serum, and centrifuged at a speed of 3000r / min for 30 minutes After the serum was separated, it was stored in a -80°C refrigerator for later use.

[0038] from F by phenol-chloroform extraction 2 Genomic DNA was extracted from the blood samples of the generation resource group, dissolved in TE, and stored at 4°C for later use. The specific method is as follows:

[0039] (1) Add 500 μL of TE buffer, 10 μL of proteinase K (10 mg / mL), 2 μL of SDS lysate to 80 μL of whole blood, shake for 10 minutes to mix well, and bathe overnight at 55 °C;

[0040] (2) Add 600 μL T...

Embodiment 2

[0049] Embodiment 2, the detection of chicken miRNA-1704 polymorphism

[0050] 1. Construction of DNA pool

[0051] 70 DNA samples with a concentration of 100ng / μL were randomly selected, and 10 μL of DNA was mixed to construct a DNA pool.

[0052] 2. Amplification primer design

[0053] Taking the homologous conserved region sequence of the genome sequence of the chicken miRNA-1704 gene in Genbank as a reference (accession number: MI0007439), Primer5.0 was used to design the sequencing PCR primer pair of chicken miRNA-1704, and the total length of the amplified fragment was 332 bp.

[0054] The sequencing primer pair sequences are as follows:

[0055] Forward primer: 5'-AGCTCTGTTGGGTTGAAGGAGTAG-3' (SEQ ID NO.1);

[0056] Reverse primer: 5'-GTGCATCTCCCACAGCTGCCATAG-3' (SEQ ID NO.2);

[0057] 3. DNA sequence of chicken miRNA-1704 cloned by PCR

[0058] Using the DNA pool as a template, carry out PCR amplification with corresponding primer pairs, and the PCR reaction system...

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Abstract

The present invention discloses single nucleotide polymorphism, a detection method and an application of chicken miRNA-1704 gene, wherein the single nucleotide polymorphism is that the site 148 in the chicken miRNA-1704 gene sequence represented by SEQ ID NO.3 is C or G, and the detection method comprises: adopting DNA of the chicken complete-genome containing the miRNA-1704 gene as a template, designing a pair of primers, amplifying, carrying out digestion on the PCR amplification product, and identifying whether the C and G single nucleotide polymorphism exists on the site 148 in the chicken miRNA-1704 gene. In addition, the detection result and the chicken growth traits are adopted to carry out correlation analysis, such that the results show that the site is significantly associated with the weights at different developmental stages; and the detection method can be used for chicken assisted selection and molecular breeding so as to rapidly establish chicken flocks with excellent genetic resources.

Description

technical field [0001] The invention relates to a single nucleotide polymorphism of chicken miRNA-1704 gene, and also relates to a detection method and application of the single nucleotide polymorphism, belonging to the field of molecular genetics. Background technique [0002] China is the country with the largest amount of poultry breeding in the world. At the same time, it has a long history of raising chickens. It is one of the countries with the earliest domesticated poultry and the richest variety resources in the world. With the rapid development of social economy and the improvement of people's living standards, the market's demand for meat quality has generally shifted from the previous quantity requirement to the pursuit of quality and flavor. Growth traits, meat quality traits and carcass traits are the most economical traits of concern to the poultry industry today. Most economic traits are quantitative traits, which are mainly determined by a large number of mi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/178
Inventor 孙桂荣王顺红康相涛李红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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