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MiRNA (micro ribonucleic acid) molecular marker miR-145 for quickly detecting survival of embryos of sows and application of miRNA molecular marker miR-145

A technology of mir-145 and molecular markers, applied in the field of molecular biology, can solve the problems of loss of profit in pig production, time-consuming and labor-intensive, waste of feed labor costs, etc.

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

AI Technical Summary

Problems solved by technology

This method is not only time-consuming and labor-intensive, but also causes a lot of waste of feed and labor costs, causing losses to the interests of pig production

Method used

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  • MiRNA (micro ribonucleic acid) molecular marker miR-145 for quickly detecting survival of embryos of sows and application of miRNA molecular marker miR-145
  • MiRNA (micro ribonucleic acid) molecular marker miR-145 for quickly detecting survival of embryos of sows and application of miRNA molecular marker miR-145
  • MiRNA (micro ribonucleic acid) molecular marker miR-145 for quickly detecting survival of embryos of sows and application of miRNA molecular marker miR-145

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Comparison of miRNA expression profiles in endometrial tissues of sows with fewer embryos survived by microarray method

[0056] 30 days after mating of Landrace sows, veterinary B-ultrasound was used to check the pregnancy status of sows, and 5 sows with less than 8 embryo implantation and 5 sows mated at the same time but with more than 8 embryo implantation sites were selected for slaughter , take endometrial samples from the implantation site, and freeze them at -70°C for testing.

[0057] RNA was extracted from endometrial tissue according to the following method.

[0058] ①Grind the endometrial tissue with liquid nitrogen, grind it into powder, transfer it to a 1.5mLEP tube, add 0.1g of 1mL Trizol solution, mix it upside down, and let it stand at room temperature for 5 minutes;

[0059] ②Add 0.2mL chloroform for every 1mL Trizol. Close the cap of the sample tube tightly, shake vigorously for 15 sec to make it fully mixed, let stand at room temperature ...

Embodiment 2

[0067] Embodiment 2 verifies the result of the chip with real-timePCR technology

[0068] 30 days after mating of Landrace sows, veterinary B-ultrasound was used to check the pregnancy status of sows, and 5 sows with less than 8 embryo implantation and 5 sows mated at the same time but with more than 8 embryo implantation sites were selected for slaughter , take endometrial samples from the implantation site, and freeze them at -70°C for testing.

[0069] RNA was extracted from endometrial tissue according to the following method.

[0070] ①Grind the endometrial tissue with liquid nitrogen, grind it into powder, transfer it to a 1.5mLEP tube, add 0.1g of 1mL Trizol solution, mix it upside down, and let it stand at room temperature for 5 minutes;

[0071] ②Add 0.2mL chloroform for every 1mL Trizol. Close the cap of the sample tube tightly, shake vigorously for 15 sec to make it fully mixed, let stand at room temperature for 5 min, and then centrifuge at 12000rmp for 15 min; ...

Embodiment 3

[0090] Example 3 Using real-timePCR technology to detect the expression level of miR-145 in the serum of sows with fewer embryonic survivors

[0091] Half an hour before breeding sows, 10ml of blood was collected in a coagulation-promoting tube, centrifuged at 2000-3000rpm for 18-20min, the supernatant was drawn into an RNase-free centrifuge tube, aliquoted in 400μl, and frozen at -80°C for testing.

[0092] Research sample selection criteria: sows were mated with veterinary B-ultrasound to check early pregnancy. The number of implanted embryos on the 30th day of pregnancy is less than 8, which is defined as the group with a small number of surviving embryos; the number of implanted embryos on the 30th day of pregnancy is more than 8, which is defined as the normal control group:

[0093] Number of serum samples collected: A total of 100 samples were collected, of which 50 were from the group with a small number of viable embryos and 50 were from the normal control group.

[...

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Abstract

The invention discloses application of a microRNA (micro ribonucleic acid)-miR-145 molecular marker to quickly detecting the survival of embryos of sows. The microRNA molecular marker in endometrial tissues of the sows with low embryo survival numbers is abnormally high in expression and can be secreted in serum via active secretion procedures of tissues, and serum miR-145 is the molecular marker with a function of quickly detecting the survival of the embryos of the sows as discovered by means of large-sample detection on the expression level of the serum miR-145 of the sows with the low embryo survival numbers. The application has the advantages that the embryo survival rates of the sows are predicted, the sows with poor embryo survival can be eliminated as soon as possible, and accordingly economic benefits can be effectively increased for pig raising enterprises.

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 rapidly detecting miRNA molecular markers miR-145 and miR-145 for the survival of sow embryos. Background technique [0002] Increasing the litter size of sows can improve the reproductive performance of sows, thereby effectively improving the economic benefits of pig farming enterprises. Embryo survival rate is one of the important factors affecting sow litter size. In pig production, how to eliminate sows with poor embryo survival as early as possible has become a difficult problem for producers. At present, the method of eliminating sows with poor embryo survival in production is mainly realized by observing the return rate and abortion rate of sows at 21 days. This method is not only time-consuming and laborious, but also causes a large amount of waste of feed and labor costs, causing losses to the interests of pig producti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113C12N15/11
CPCC12Q1/6888C12Q2600/158C12Q2600/178
Inventor 吴德周盼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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