Unlock instant, AI-driven research and patent intelligence for your innovation.

Primer sets and detection kits for the detection of components derived from horses, donkeys, mules, and donkey mules

A technology for detecting kits and source components, which is applied in the direction of recombinant DNA technology, biochemical equipment and methods, measurement/testing of microorganisms, etc., can solve the problems of cumbersome and time-consuming components, and achieve short time-consuming and high workload Small, cost-saving effect

Active Publication Date: 2022-06-21
INNER MONGOLIA AGRICULTURAL UNIVERSITY
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] To this end, the embodiment of the present invention provides a primer set and detection kit for detecting components derived from horses, donkeys, horse mules and donkey mules, so as to solve the problem of horses, donkeys, horse mules and donkey mules detected in the prior art. Source ingredients are cumbersome and time-consuming

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Primer sets and detection kits for the detection of components derived from horses, donkeys, mules, and donkey mules
  • Primer sets and detection kits for the detection of components derived from horses, donkeys, mules, and donkey mules
  • Primer sets and detection kits for the detection of components derived from horses, donkeys, mules, and donkey mules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] In the embodiment of the present invention, there is a small fragment deletion between the homologous genes of horse and donkey found in the autosomal chromosome, and a first specific primer pair is designed at both ends of the deletion region. In order to increase the difference, when designing primers, the Base substitution is used to increase the specificity of the primer pair, and the nucleotide sequence of the first specific primer pair is the upstream primer shown in SEQ ID NO:1 and the downstream primer shown in SEQ ID NO:2. The horse's homolog CNGB3 cyclic nucleotide gated channelsubunit beta 3, specifically, horse [Equus caballus(horse)] Gene ID: 100055470, LocationChromosome 9, NC_009152.3(2298952..2413636); donkey [Equus asinus(ass) ]Gene ID: 106829842 Location NW_014637333.1(39847..171617). The nucleotide sequence of the donkey homologous gene amplified by the first specific primer pair is shown in SEQ ID NO: 7, and the nucleotide sequence of the amplified h...

Embodiment 2

[0034] In the embodiment of the present invention, the mitochondrial genes of horses and donkeys were used to identify and distinguish between mules and donkeys, and the full-length mitochondrial mitochondrial DNA (mtDNA) of horses and donkeys were downloaded from NCBI, wherein, Horse Horse: ACCESSION NC_001640LOCUS NC_001640 16660bp ; Donkey Dokey: ACCESSION NC_001788LOCUS NC_00178816670bp, use BioEdit software to compare, find the DNA segment that is very different in mitochondrial genes, and use this DNA segment to design nucleotide sequences such as SEQ ID NO: 3 and SEQ ID NO: The second specific primer pair shown in 4. In order to increase the difference, when designing the primers, base substitution was performed to increase the specificity of the primer pair, and the nucleotide sequence of the amplified PCR product is shown in SEQ ID NO: 9. During the extraction of the template for sample PCR, the extraction of blood DNA was performed using the TaKaRa MiniBEST Universal...

Embodiment 3

[0038] In the embodiment of the present invention, SRY (mammalian early gonads have the potential for bidirectional differentiation. In the process of sex determination, the undifferentiated gonads are finally determined according to whether the somatic cells express the SRY (sex-determining region of the Y chromosome) gene or not). It develops into ovarian or testis gene, and the unique gene on the Y chromosome is used to design the third specific primer pair. In order to increase the difference, when designing primers, base substitution is carried out to increase the specificity of the primer pair. Its nucleotides The sequences are shown in SEQ ID NO: 5 and SEQ ID NO: 6, and the nucleotide sequence of the amplified PCR product is shown in SEQ ID NO: 10.

[0039] Among them, the PCR amplification system is: using Green Taq Mix (Vazyme, p131), 20ul reaction system, containing Green Taq Mix 10ul, DNA template 2.4ul, upstream primer 0.5ul, downstream primer 0.5ul, ddH 2 O 6.6ul....

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The embodiment of the present invention discloses a primer set and a detection kit for detecting components derived from horses, donkeys, horse mules and donkey mules. The primer set includes nucleotide sequences such as SEQ ID NO: 1 and SEQ ID NO: 2 The first pair of specific primers shown, the second specific primer pair shown in SEQ ID NO: 3 and SEQ ID NO: 4, the primer set also includes the A third pair of sex-specific primers for horses, donkeys, mules and donkey mules, the nucleotide sequences of the third pair of specific primers are shown in SEQ ID NO:5 and SEQ ID NO:6. And a multiplex PCR detection kit containing the primer set. The primer set and detection kit for detecting components derived from horses, donkeys, mules and donkeys according to the embodiments of the present invention, in the process of detecting components derived from horses, donkeys, mules and donkeys, it takes a short time , the workload is small, the detection result is accurate, and the detection cost is saved.

Description

technical field [0001] The embodiments of the present invention relate to the technical field of biological detection, in particular to a primer set and a detection kit for detecting components derived from horses, donkeys, mules and donkey mules. Background technique [0002] At present, horses and donkeys are homologous species with a high degree of similarity in genomic information, and the two can be hybridized to produce mules. The similarity between horse mules and donkeys and mules is particularly high, and mistakes are often made by visual observation; It is difficult to distinguish the tissue samples of horses, mules, donkeys and mules by general methods, especially the sources of raw materials from horses, donkeys, mules, donkeys and mules, such as donkey-hide gelatin and other products, which are made of donkey hides after boiling. , concentrated and made, it has the functions of improving sleep, strengthening the brain and improving intelligence. However, in thi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/16
Inventor 格日乐其木格王希生张心壮神英超纳日嘎芒来
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY