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Applications of microRNA in swine intramuscular fat detection

An intramuscular fat and internal reference technology, which can be used in DNA/RNA fragments, recombinant DNA technology, and microbial determination/inspection, etc., and can solve problems such as constant expression

Inactive Publication Date: 2014-08-13
HUAZHONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no ideal internal reference gene that can be completely used in any experimental conditions, and the expression is constant. The applicant should choose a suitable and stable internal reference gene for calibration and standardization according to the purpose of the experiment and the type of sample, in order to obtain reliable The experimental results

Method used

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  • Applications of microRNA in swine intramuscular fat detection
  • Applications of microRNA in swine intramuscular fat detection
  • Applications of microRNA in swine intramuscular fat detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] 1. Construction of pig family and determination of pig intramuscular fat

[0062] The experimental pigs used for analysis are purebred large white pigs (conventional breeds, the FCR project test group established by the Animal Genetics, Breeding and Reproduction Molecular Biology Laboratory of Huazhong Agricultural University and Pfizer Pharmaceutical Co., Ltd. according to the conventional method. The experimental pig farm of Huazhong Agricultural University, a total of 236 pigs, large white pigs are commonly used breeds from foreign blood pigs), slaughtered when the body weight reached 80-100kg, the samples were collected in 21 batches, and the traits were measured after slaughter, including those used for correlation analysis Body length data, including: carcass length (abbreviated as CL), carcass oblique length (abbreviated as CDL), rib number (abbreviated as RN), intramuscular fat (abbreviated as IMF), etc.

[0063] Intramuscular fat was measured on the longissimus...

Embodiment 2

[0070] Differential expression detection of porcine U6, 18S rRNA, and mir133a3p genes in individuals with different intramuscular fat contents

[0071] 1. Select individuals and extract pig genome RNA.

[0072] The samples are purebred large white pigs (the same source as above), with a body weight of about 90Kg.

[0073] Three individuals with high and low intramuscular fat were selected from the population in the above-mentioned Example 1, and the total RNA of the longissimus dorsi muscle was extracted (the preparation method of total RNA is a conventional method, and the specific steps are as follows). The intramuscular fat content of the longissimus dorsi muscle was measured for each individual, and the statistical results are shown in Table 1.

[0074] Total RNA was extracted using Invitrogen Reagent kit extraction (operate according to the instructions of the kit), the specific steps are as follows:

[0075] (1) Take out the preserved longissimus dorsi muscle tissue ...

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Abstract

The invention belongs to the technical field of livestock molecular marker preparation and application, and particularly relates to applications of a miRNA marker and a specific primer thereof relevant to swine intramuscular fat. The miRNA marker gene is swine-based microRNA-133a3p gene, the expression quantity difference of the microRNA in swine intramuscular fat can be detected by utilizing the gene as a reference gene through adopting a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) method, the results show that the marker of the Ssc-miR133a3p gene prepared by cloning is high in expression quantity, strong in stability, and superior to the primer of U6 and 18S rRNA genes in repeatability.

Description

technical field [0001] The invention belongs to the technical field of preparation and application of livestock molecular markers, and specifically relates to a miRNA marker related to pig intramuscular fat and the application of specific primers. The miRNA marker gene is based on the pig microRNA-133a3p gene. The gene was used as an internal reference gene to detect the expression of microRNA in pig intramuscular fat by real-time fluorescent quantitative PCR method. Background technique [0002] With the improvement of people's quality of life, consumers have correspondingly increased quality requirements for pork, such as the taste, juiciness and tenderness of pork. Intramuscular fat (IMF) or marbling plays a vital role in pork quality (Tan Lin and Jiang Hailong, 2010). IMF is mainly distributed between muscle bundles and muscle fibers, and it melts out from between muscle fibers to make the meat tender and juicy. Studies have shown that IMF is positively correlated with...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/166C12Q2600/178
Inventor 曹建华刘慧莹赵书红李长春李新云朱猛进余梅
Owner HUAZHONG AGRICULTURAL UNIVERSITY