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Binary pig gene identification method

An identification method and gene technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of cumbersome detection process and low accuracy of detection results, and achieve the effect of simple detection operation and improved accuracy

Pending Publication Date: 2021-10-12
北京康普森农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After comparing the gene sequences, it was found that all Landrace and Large White MC1R genes do not have the G1554A mutation, and all Duroc MC1R genes have the G1554A mutation. From this, it can be seen that there is no G1554A mutation in the binary pig (Landrace X Large White cross), so Designing binary pig detection products based on this mutation site can help companies recover losses, which is of great significance. Existing detection products have the disadvantages of low accuracy of detection results and cumbersome detection process

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Embodiment 1

[0021] 1.1 Sample selection

[0022] 163 sows were selected, of which 20 binary sows were pure binary sows obtained by crossing Landrace pigs and Large White pigs, and 77 three-yuan sows were non-binary sows obtained by crossing Duroc pigs and Dongbeimin pigs , 66 binary sows are non-pure binary sows obtained by crossing Duroc pigs and Landrace pigs.

[0023] 1.2 Primer design

[0024] After sequence comparison, it was found that there was no G1554A mutation in the MC1R gene of Landrace and Large White, and the first forward primer, the second forward primer and the universal reverse primer were designed based on their genome sequences.

[0025] The nucleotide sequence of the first forward primer is shown in SEQ ID NO:1, the first forward primer sequence includes a specific fluorescent sequence FAM, the specific fluorescent sequence FAM is combined with a FAM fluorescent marker, and the nucleotide sequence of the specific fluorescent sequence FAM As shown in SEQID NO:4;

[...

Embodiment 2

[0033] 2.1 Sample selection

[0034] 163 sows were selected, of which 114 binary sows were pure binary sows obtained by crossing Landrace pigs and Large White pigs, and 5 three-yuan sows were non-binary sows obtained by crossing Duroc pigs and Dongbeimin pigs , 44 binary sows are non-pure binary sows obtained by crossing Duroc pigs and Landrace pigs.

[0035] 2.2 Primer design

[0036] After sequence comparison, it was found that there was no G1554A mutation in the MC1R gene of Landrace and Large White, and the first forward primer, the second forward primer and the universal reverse primer were designed based on their genome sequences.

[0037] The nucleotide sequence of the first forward primer is shown in SEQ ID NO:1, the first forward primer sequence includes a specific fluorescent sequence FAM, and the nucleotide sequence of the specific fluorescent sequence FAM is shown in SEQ ID NO:4;

[0038] The nucleotide sequence of the second forward primer is shown in SEQ ID NO:...

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Abstract

The invention relates to the technical field of gene detection, in particular to a binary pig gene identification method which comprises the following steps: taking a first allele and a second allele of an MC1R gene of a to-be-detected pig as templates, carrying out PCR amplification by using a first forward primer, a second forward primer and a universal reverse primer, and carrying out fluorescence detection on an amplification product by adopting a fluorescence detection instrument, and performing SNP typing on the MC1R gene of the to-be-detected pig according to the fluorescence type of the amplification product. According to the invention, two forward primers and a universal reverse primer are designed, gene identification is carried out on a binary pig (Changbai X big white hybrid) by utilizing the characteristics that the MC1R genes of Changbai and big white pigs do not have G1554A mutation and the Duroc MC1R gene has G1554A mutation, and the accuracy of a detection result is improved; according to the invention, a fluorescent probe technology is adopted, a specific fluorescent sequence FAM and a specific fluorescent sequence HEX are respectively connected to one ends of the two forward primers, and a PCR amplification product is subjected to fluorescence detection through detection, so that gene identification can be carried out on the binary pig, and the detection operation is simpler.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a binary pig gene identification method. Background technique [0002] Binary pigs are hybrids between two breeds. The commonly used crossbreeding method in the market is Large White and Landrace pigs. As the situation of Sanyuan pigs replacing Binary pigs in the market is becoming more and more serious, in order to avoid unnecessary losses for farmers, The detection and identification of binary pigs is of great significance. After comparing the gene sequences, it was found that all Landrace and Large White MC1R genes do not have the G1554A mutation, and all Duroc MC1R genes have the G1554A mutation. From this, it can be seen that there is no G1554A mutation in the binary pig (Landrace X Large White cross), so Designing detection products for binary pigs based on this mutation site can help companies recover losses, which is of great significance. Existing detection produ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12Q1/6888
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2563/107
Inventor 刘继强
Owner 北京康普森农业科技有限公司