Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Fluorescent automated detection method of pathogenic sites of Arachnomelia syndrome (AS) in Simmental cattle

A technology for automatic detection of Simmental cattle, applied in biochemical equipment and methods, fluorescence/phosphorescence, microbial measurement/inspection, etc., to achieve stable results and low cost

Inactive Publication Date: 2012-07-04
CHINA AGRI UNIV
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there is no rapid and accurate detection method for the AS pathogenic locus in Simmental cattle that is suitable for commercial operation

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
  • Fluorescent automated detection method of pathogenic sites of Arachnomelia syndrome (AS) in Simmental cattle
  • Fluorescent automated detection method of pathogenic sites of Arachnomelia syndrome (AS) in Simmental cattle
  • Fluorescent automated detection method of pathogenic sites of Arachnomelia syndrome (AS) in Simmental cattle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Preparation of test materials and extraction of DNA

[0034] The test materials used in this study came from two parts: the anticoagulant blood samples and ear tissue samples of the mating cows and hybrids from the family of 4 German Simmental bulls; and the frozen semen of the bulls of 11 cattle breeds or strains.

[0035] 4 German Simmental bull pedigrees (Table 1), including 80 and mating cows and 106 offspring, from Anshan Hengli Dairy Farm (Anshan Hengli) in China and Sheltala Sanhe cattle breeding farm in Hailar, Inner Mongolia (Hailal Sheltala). Among the four families, the cow breeds are Chinese Holstein and Chinese Sanhe cattle, and the offspring are all hybrid cattle (Holstein × German Simmental crossbreed, Sanhe cattle × German Simmental crossbreed). The 4 German Simmental cows are ROMEL, BOSSAG, RIFURT, HIRMER (German Simmental genealogy query website: http: / / www.zar.at / article / archive / 18981). ROMEL is a known carrier of the AS pathogenic gene, and th...

Embodiment 3A

[0060] Example 3 Direct sequencing analysis of AS pathogenic mutations

[0061] In this study, primers were designed for the c.del1224_1225CA mutation of MOCS1 gene, and 4 German Simmental bulls such as ROMEL and 31 offspring of ROMEL’s pedigree records were selected to carry out PCR amplification of the mutation fragments. Perform PCR product sequencing. The sequencing primers are shown in Table 5. The PCR system is the same as in Example 2, and the PCR reaction is as shown in Figure 4 Shown. PCR was detected by 2% gel, and the PCR product was purified and Sanger sequenced at BGI.

[0062] Table 5 Primers for direct sequencing of MOCS1 gene mutation sites

[0063]

[0064] The generation (Table 6) is the heterozygous genotype carrying the mutation, and other individuals are wild-type. ROMEL and its 2 progeny sequencing peaks are as shown Image 6 As shown, ROMEL is heterozygous at the mutation site, and its two progeny 94254 and 9129x are heterozygous and wild homozygous at the ...

Embodiment 4A

[0069] Example 4 AS Pathogenic Mutation Fluorescence Automated Detection Method

[0070] 1. Method

[0071] In this study, the MOCS1 gene c.del1224_1225CA mutation is a 2bp deletion mutation. Due to the high sensitivity of fluorescent primer PCR detection method, it can accurately detect the length polymorphism of 1bp. Therefore, this study designed fluorescently labeled primers for the mutation site. The genotype was detected by capillary electrophoresis after PCR amplification. Because there may be systematic errors between different plates in capillary electrophoresis detection, there are 3 identical individuals in each plate in the experiment to correct the genotyping errors between the plates; the intra-plate errors are corrected using the molecular weight standard GS500LIZ.

[0072] The 5'end of the upstream primer is labeled with a FAM fluorophore. The PCR system and reaction procedure are the same as in Example 2, and the reaction procedure is as follows Figure 4 Shown.

[...

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 invention relates to a fluorescent automated detection and analysis method of pathogenic sites of Arachnomelia syndrome (AS) in Simmental cattle. The nucleotide sequence of a primer for PCR (polymerase chain reaction) amplification is as follows: 5'-CTGAGTCTCCTCTTCTGTTTTCA-3',5'-GTTGGCATCTGAGTCCAGGT-3'. By using the fluorescent labeling primer PCR amplification and electrophoresis typing technology, the kit for typing pathogenic sites of AS in Simmental cattle has the advantages of stable result and low cost, and is accurate and reliable, thereby accurately typing the pathogenic sites of AS in Simmental cattle.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a PCR detection method with fluorescent marker primers at the pathogenic site of Simmental spider leg syndrome (AS). Background technique [0002] The reason for the recessive mode of genetic defects in livestock may have something to do with the production methods of livestock, especially dairy cows. The application of artificial insemination (AI) and semen cryopreservation techniques (Semen Cryopreservation Techniques) in the production and breeding of dairy cows has laid a solid foundation for the establishment of an AI breeding system for dairy cows. Because AI technology can take on more female breeding tasks, bulls can be widely used in groups to obtain a large number of offspring, so that high-yielding genes can be rapidly spread in the group. Semen cryopreservation technology allows excellent breeding bulls to realize the exchange of genetic material on a global scale...

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
IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 王雅春焦士会初芹俞英张胜利孙东晓张毅张沅
Owner CHINA AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com