Fluorescently-labeled gene multiplex amplification method for identifying sources of goat, sheep, pig and duck meat simultaneously

A fluorescent labeling, sheep technology, used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of no simultaneous detection, avoid cross-interference, high detection sensitivity, and improve detection efficiency. Effect

Active Publication Date: 2013-11-20
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
View PDF8 Cites 44 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the improvement of adulteration means, the combination of adulterated species is becoming more and more diverse, among which pork, duck and chicken are the most common in mutton, but there is no simultaneous detection of goat, sheep, pig and duck. The method for growing meat, the present invention uses mitochondrial 16S rRNA and cyt b gene as the target gene, uses two pairs of primers to car

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
  • Fluorescently-labeled gene multiplex amplification method for identifying sources of goat, sheep, pig and duck meat simultaneously
  • Fluorescently-labeled gene multiplex amplification method for identifying sources of goat, sheep, pig and duck meat simultaneously
  • Fluorescently-labeled gene multiplex amplification method for identifying sources of goat, sheep, pig and duck meat simultaneously

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0048] Primer design and fluorescent labeling: According to the mitochondrial DNA of goat (accession number M55541.1), sheep (accession number NC_001941.1HM236185.1), duck (accession number X60825.1) and pig (accession number AY920914.1) published in GenBank 16s rRNA gene sequence, design a pair of universal primers, see the binding position of the primers in the mitochondrial genome figure 1 ; According to the cyt b sequence of the above species, design a pair of universal primers as internal reference. Then label the 5' end of the forward primer in each pair of primers with a 6-FAM fluorescent molecule, that is, the forward primer SIM-F of 16S rRNA: 5'-FAM-AAGACGAGAAGACCCTTGGACTTTA-3', the reverse primer SIM-R of 16S rRNA: 5'-GATTGCGCTGTTATCCCTAGGGTA-3'; forward primer mcb-F of cyt b gene: 5'-FAM-TACCATGAGGACAAATATCATTC TG-3', reverse primer mcb-R of cyt b gene: 5'-CCTCCTAGTTTGTTAGGGAT TGATCG-3'.

[0049] See Table 1 for the sequences of universal primers for mitochondrial ...

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 provides a fluorescently-labeled gene multiplex amplification technology for detecting sources of goat, sheep, pig and duck meat simultaneously. A method comprises the steps that mitochondria 16S rRNA (ribosomal Robonucleic Acid) and Cyt (Cytosine) b genes of a goat, sheep, a pig and a duck serve as target genes, and are subjected to PCR (Polymerase Chain Reaction) multiplex amplification by two pairs of fluorescently-labeled primers; PCR amplification products are detected by a 3730xl full automatic gene analysis meter; and species identification is performed according to 16S rRNA gene amplified fragment length polymorphism. The method fills in a gap of a detection method of the four kinds of meat.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a fluorescent marker gene composite amplification method capable of simultaneously identifying four kinds of meat origins such as goat, sheep, pig and duck. Background technique [0002] Food is food to safety first. Meat safety is a major issue related to people's livelihood. Mutton has less fat, rich protein, high calcium, potassium and VB1, low cholesterol content, high nutritional value and food tonic value, and has been favored by people all the time. In recent years, it has been continuously reported that criminals have mixed a large amount of pork, duck and chicken with a small amount of beef and mutton fat and leftovers to sell as mutton, earning high profits. It has become a "hidden rule" in the circle to make lamb skewers with pork and duck meat with flavor and pigment. Reports about fake mutton continue to appear in the newspapers, which not only brings...

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): C12Q1/68C12N15/11
Inventor 步迅张全芳刘艳艳
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap