MicroRNA (micro ribonucleic acid) molecular marker miR-23a for quickly detecting quality of oocytes of sows and application of microRNA molecular marker miR-23a

A mir-23a, oocyte technology, applied in DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of sow injury, non-instructive production, staying in the laboratory, etc.

Active Publication Date: 2015-12-02
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current quality detection methods for sow oocytes are still in the laboratory st

Method used

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  • MicroRNA (micro ribonucleic acid) molecular marker miR-23a for quickly detecting quality of oocytes of sows and application of microRNA molecular marker miR-23a
  • MicroRNA (micro ribonucleic acid) molecular marker miR-23a for quickly detecting quality of oocytes of sows and application of microRNA molecular marker miR-23a
  • MicroRNA (micro ribonucleic acid) molecular marker miR-23a for quickly detecting quality of oocytes of sows and application of microRNA molecular marker miR-23a

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Microarray method to compare the differences in miRNA expression profiles in cumulus oocytes with low developmental potential

[0058] The commercial hog ovaries were collected from the slaughterhouse, and the ovaries were transported back to the laboratory as soon as possible in sterile saline at 37 °C. Use a sterile syringe with a 20-gauge needle to extract follicles with a diameter of 2-6mm on the surface of the ovary, and slowly inject the extracted follicle fluid into a high-pressure sterilized centrifuge tube to allow the oocytes to settle naturally for 10 minutes, and then use a pipette to slowly Aspirate the follicle supernatant slowly, and transfer the precipitate after removing the supernatant to a 60mm culture dish containing PBS buffer. Divide the cumulus oocytes into two types with a mouth pipette under a stereomicroscope and transfer them to 1ml centrifuge tubes, and store them at -70°C for testing. Classification criteria: low developmental pot...

Embodiment 2

[0069] Embodiment 2 verifies the result of the chip with real-timePCR technology

[0070] The commercial hog ovaries were collected from the slaughterhouse, and the ovaries were transported back to the laboratory as soon as possible in sterile saline at 37 °C. Use a sterile syringe with a 20-gauge needle to extract follicles with a diameter of 2-6mm on the surface of the ovary, and slowly inject the extracted follicle fluid into a high-pressure sterilized centrifuge tube to allow the oocytes to settle naturally for 10 minutes, and then use a pipette to slowly Aspirate the follicle supernatant slowly, and transfer the precipitate after removing the supernatant to a 60mm culture dish containing PBS buffer. Divide the cumulus oocytes into two types with a mouth pipette under a stereomicroscope and transfer them to 1ml centrifuge tubes, and store them at -70°C for testing. Classification criteria: low developmental potential cumulus oocytes: oocytes with uniform cytoplasm, only a...

Embodiment 3

[0092] Example 3 Using real-timePCR technology to detect the expression level of miR-23a in the serum of sows with poor oocyte quality

[0093] Serum sample collection: Collect 10ml of blood on an empty stomach the next morning when sows appear standing reflex, centrifuge at 2000-3000rpm for 18-20min, draw the supernatant into RNase-free centrifuge tubes, aliquot 400μl, and freeze at -80℃ until Measurement.

[0094] Research sample selection criteria: Landrace sows with less than 5 litters in two consecutive parities, body condition score (2.5-3.5), and normal and healthy food intake are defined as the poor oocyte quality group; Healthy Landrace sows with the same parity, two consecutive parities with 10-13 litters, body condition score (2.5-3.5), and normal feed intake were defined as the normal control group. A total of 100 samples were collected, 50 of which were from the low litter size group and 50 from the normal control group.

[0095] Extraction of serum microRNA

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Abstract

The invention discloses application of a microRNA (micro ribonucleic acid)-miR-23a molecular marker miR-23a to quickly detecting the quality of oocytes of sows. The miRNA molecular marker miR-23a in the cumulus oocytes with low potentiality development is abnormally high in expression and can be secreted in serum via active secretion procedures of tissues, and serum miR-23a is the molecular marker with a function of quickly detecting the quality of the oocytes of the sows as discovered by large-sample detection on the expression level of the serum miR-23a of the sows with low production performance. The application has the advantages that the reproductive potential of the sows can be predicted by means of detecting the quality of the oocytes, so that the sows with low production potential can be eliminated as early as possible, the feed and labor costs can be greatly reduced, and the application is beneficial to vigorous development of the pig industry in China.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a primer, a kit and a detection method for miRNA molecular markers miR-23a and amiR-23a for rapidly detecting the quality of sow oocytes. Background technique [0002] The economic benefits of pig farms are fundamentally related to the reproductive rate of sows. The reproductive performance of sows is an important economic indicator in pig production. In pig production, in addition to giving full play to and utilizing the reproductive performance of sows, it is also necessary to eliminate sows with low production performance in time to maximize economic benefits. How to eliminate sows with poor production performance as soon as possible has become a problem that plagues pig production. The study found that sow oocyte quality is positively correlated with production performance. Therefore, testing the quality of sow oocytes is the most accurate and effective way to eval...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/113C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/178
Inventor 吴德晋超方正锋车炼强徐盛玉林燕冯斌李建陈玲赵希仑
Owner SICHUAN AGRI UNIV
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