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Primers, kits and methods for paternity testing of cervidae animals

A parent-child identification and kit technology, which is applied in the field of primers and kits for parent-child identification of cervidae, can solve the problems of poor identification effect and achieve clear bands, simple identification method and reliable identification results

Active Publication Date: 2020-08-28
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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AI Technical Summary

Problems solved by technology

Studies have shown that only 56% of bovine microsatellite primers can be applied to sheep, and only 42% of the microsatellite sites are polymorphic in sheep, and the identification effect is poor

Method used

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  • Primers, kits and methods for paternity testing of cervidae animals
  • Primers, kits and methods for paternity testing of cervidae animals
  • Primers, kits and methods for paternity testing of cervidae animals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0044] 1. Primer Synthesis

[0045] Synthesized 10 pairs of microsatellite primers for paternity identification of deer, with different fluorescent markers (FAM, HEX, TAMRA) at the 5' ends of the primers. The specific information is as follows:

[0046] Table 1 10 pairs of microsatellite primers and their groupings for paternity testing of deer

[0047]

[0048]

[0049] 2. Genome extraction of the sika deer to be tested

[0050] Take 300ul of sika deer blood, use the Biotake blood extraction kit to extract the sika deer genome, use 1% agarose gel electrophoresis to detect the integrity of the DNA, measure the DNA concentration and purity with a microplate reader, and store it at -20°C for later use.

[0051] 3. Using the sika deer genome to be detected as a template, the above 10 microsatellite loci were amplified by PCR.

[0052] The PCR amplification system and conditions are shown in Table 2 and Table 3.

[0053] Table 2 PCR reaction system

[0054]

[0055] ...

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Abstract

The invention provides a primer, a kit and a method for paternity identification of deer animals; the primer is selected from at least one primer pair of the following: SEQ ID NO: 1-20. This set of primers has a total of 10 pairs, and the amplified bands are clear and highly polymorphic, and the cumulative non-father exclusion rate is as high as 99.99%, which can be applied to the practice of deer paternity testing.

Description

technical field [0001] The invention relates to the field of molecular genotyping, in particular to a primer, a kit and a method for paternity identification of deer animals. Background technique [0002] Microsatellite is a short tandem repeat sequence (2-6bp) in the genome, which shows different alleles due to different repeat numbers. Compared with other identification methods, paternity testing with microsatellite markers has many incomparable advantages. Among them, the most notable advantage is his co-dominant genetic characteristics, which allows homozygous and heterozygous genotypes to be distinguished, greatly increasing the accuracy of paternity testing. In addition, microsatellite markers have high polymorphism, that is to say, there are multiple alleles at the same locus. Since non-parent-child individuals are unlikely to share the same allele, a small number of high polymorphisms is used. Morphological microsatellite markers can identify the parent-child relat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2525/151
Inventor 王桂武杨万云
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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