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Single nucleotide polymorphism (SNP) of ADD1 gene of cattle and detection method thereof

A single nucleotide polymorphism and detection method technology, which is applied in the application field of cattle breeding, can solve problems such as inconsistent research results, and achieve the effects of easy popularization and application, high accuracy and low cost

Inactive Publication Date: 2012-07-04
XUZHOU NORMAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

However, the research results on the relationship between ADD1 gene variation and hypertension are quite inconsistent.
The research on ADD1 gene and growth and development traits has not been reported yet. Based on the research results of actin, the function of adducin (cell membrane skeleton protein) will be further explored

Method used

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  • Single nucleotide polymorphism (SNP) of ADD1 gene of cattle and detection method thereof
  • Single nucleotide polymorphism (SNP) of ADD1 gene of cattle and detection method thereof
  • Single nucleotide polymorphism (SNP) of ADD1 gene of cattle and detection method thereof

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Embodiment Construction

[0033] 1. Collection and processing of blood samples

[0034] Take 10 mL of blood sample, add 500 μL of 0.5 mol / L EDTA for anticoagulation, slowly invert it 3 times, put it in an ice box, and store it at -80°C for later use.

[0035] A total of 555 cattle blood samples were used in this study, specifically:

[0036] 1) 374 blood samples of Jiaxian Red Cattle were collected from Jiaxian Red Cattle Breeding Area, Pingdingshan City, Henan Province;

[0037] 2) 116 blood samples from Qinchuan cattle were collected from the Yellow Cattle Breeding Farm in Nanyang City, Henan Province;

[0038] 3) 65 blood samples from Luxi cattle were collected from Shandong cattle farm.

[0039] 2. Extraction of genomic DNA from blood samples

[0040] (1) Thaw the frozen blood sample at room temperature, transfer 500μL to a 1.5mL Eppendorf tube, add an equal volume of PBS buffer, mix well, centrifuge at 12000r / min for 10min (4℃), discard the supernatant, repeat the above steps to the supernatant ...

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Abstract

The invention discloses a method for detecting the single nucleotide polymorphism (SNP) of an ADD1 gene of cattle, which comprises the following steps: through taking a genomic DNA of cattle containing an ADD1 gene as a template and taking a primer mix P as a primer, the ADD1 gene of cattle is amplified through PCR (polymerase chain reaction), then the segment size of the obtained product is amplified through AGE (agarose gel electrophoresis) detection; and then, the detection on SSCP (single strand conformation polymorphism) is performed. The polymorphism of the gene comprises a base polymorphism that the 70027th bit in an ADD1 gene sequence table SEQID No.8 of cattle is G or A, and a base polymorphism that the 70051th bit in the ADD1 gene sequence table SEQID No.8 of cattle is G or A. The analysis on the correlation between the SSCP polymorphism and the production performance shows that: a certain correlation exits between the polymorphic site and the production performance, therefore, the weight of a Qinchuan cattle (genotype: BB) is significantly higher than that of a cattle (genotype: AA). The method is used for the auxiliary selection and molecular breeding of cattle throughscreening and detecting molecular genetic markers closely relating to the growth properties of cattle on a DNA level, therefore, the method plays a vital role in improving the production performance of cattle.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the screening and detection of single nucleotide polymorphism (SNP) of cattle ADD1 gene and its application as an auxiliary selection marker in cattle breeding. Background technique [0002] Gene polymorphism refers to the difference in genome sequence between different species or different individuals in the same species. These differences are caused by nucleotide changes in DNA alleles in chromosomes, which include base substitution, insertion, deletion, and repeated sequence copies number of changes. [0003] Single nucleotide polymorphism refers to the polymorphism caused by the replacement of a single nucleotide (A / T / C / G) in the genomic DNA sequence, mainly caused by the conversion or transversion of a single base. SNPs with transformational variants accounted for about 2 / 3, and several other SNPs were at similar levels. The cytosine of the CpG dinucleotide is the most mutated s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 陈宏钱丽娜张春雷房兴堂
Owner XUZHOU NORMAL UNIVERSITY
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