Fluorescence quantitative PCR detection primer and method for type and composition of yak muscle fibers

A technique for muscle fiber type and fluorescence quantification, which is applied to the detection primers and detection fields of fluorescence quantitative PCR composed of yak muscle fiber types, and achieves the effects of low cost, high efficiency and small workload.

Active Publication Date: 2015-11-11
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Claims
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Problems solved by technology

At present, there are relatively few studies on the types of yak muscle fibers, and no effective detection method for the composition of yak muscle fibers has been established.

Method used

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  • Fluorescence quantitative PCR detection primer and method for type and composition of yak muscle fibers
  • Fluorescence quantitative PCR detection primer and method for type and composition of yak muscle fibers
  • Fluorescence quantitative PCR detection primer and method for type and composition of yak muscle fibers

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Embodiment 1

[0040] 1. Sample collection:

[0041] The longissimus dorsi muscle tissue of yaks was collected quickly after slaughter and stored in liquid nitrogen.

[0042] 2. Reagents:

[0043] RNAsimple total RNA extraction kit: Tiangen Biochemical Technology (Beijing) Co., Ltd.;

[0044] PrimeScript? RTreagentKitwithgDNAEraser: Takara Corporation;

[0045] SYBR?PremixExTaq?II (PerfectRealTime): Takara Company;

[0046] Primer synthesis: American life company.

[0047] 3. Main instruments and equipment:

[0048] PCR instrument: Bio-rad company;

[0049] CFX-96 fluorescent quantitative PCR instrument: Bio-rad company;

[0050] Nanodrop2000 Ultra Micro Spectrophotometer: ThermoScientific Company.

[0051] 4. Detection method:

[0052] a) Design and synthesis of primers:

[0053] According to the cDNA sequence differences of different subtypes of yak MyHC gene, primers were designed using Primer Premier5.0 and Oligo6.0 software. The company synthesizes TaqMan probes. The sequences ...

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Abstract

The invention discloses a fluorescence quantitative PCR detection method for type and composition of yak muscle fibers. According to the fluorescence quantitative PCR detection method, yak muscle tissues are taken for total RNA extraction, the RNA purity and concentration are detected, the total RNA is subjected to inverse transcription to generate cDNA, then quality detection is carried out by taking GAPDH genes of yak as internal genes, and then a special detection primer is adopted for carrying out fluorescence quantitative PCR reaction, meanwhile, the GAPDH genes of yak are taken as reference genes for statistic analysis. The detection method and the detection primer provided by the invention are more accurate and more reliable than histochemistry evaluation; with the molecule quantitative classification method, accurate quantitative and analysis can be carried out on the type composition of the various muscle fibers in muscles at different parts of the yak, so that a technological base is laid for further study on the relevance between the muscle fibers and the meat quality trait of the yak, and meanwhile, compared with other methods, the fluorescence quantitative PCR detection method has the advantages of low workload and low cost, and high efficiency.

Description

technical field [0001] The invention relates to the field of detection primers and detection methods for muscle fiber types, in particular to fluorescent quantitative PCR detection primers and detection methods for yak muscle fiber types. Background technique [0002] The yak is a breed of cattle distributed in the center of the Qinghai-Tibet Plateau and its adjacent alpine and sub-alpine regions. It provides herders with milk, meat, hair, labor, fuel and other production and living materials. It has the reputation of "the boat of the plateau". Yaks eat natural pastures all year round. Their meat has high protein content, low fat content, and is rich in unsaturated fatty acids and special flavor substances. It is a green food that meets the requirements of modern life (Yan Ping, 2012). It is a research hotspot in the field of yak genetics and breeding. [0003] Muscle fiber is the main component of skeletal muscle. The difference in muscle fiber type will lead to the differ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/158
Inventor 吴晓云褚敏梁春年郭宪阎萍
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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