PCR amplification identification method and special primers based on 16s rRNA quail meat components
An identification method, quail meat technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the violation of the legitimate interests of producers and consumers, the label does not fully match the variety, animal origin Food adulteration and other issues, to achieve the effect of high practical value, low cost and fast speed
Active Publication Date: 2020-08-21
JIANGSU INST OF POULTRY SCI
View PDF1 Cites 0 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
[0002] In recent years, various food safety incidents have occurred frequently, such as adulteration of animal-derived food, shoddy quality, and the substitution of cheap and low-quality livestock and poultry meat for high-priced high-quality livestock and poultry meat, which seriously disrupted the market order and harmed producers and consumers. The legitimate interests of consumers, while bringing serious food safety risks
In the West, a test and analysis of nearly a thousand kinds of meat products showed that nearly 20% of the products did not completely match the label and the variety
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 moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment Construction
[0026] DNA extraction - phenol imitation method
[0027] (1) Take 25 mg of raw / cooked quail meat sample, cut it into pieces, add 500 μl of cell lysate (0.5% sodium dodecyl sulfate (SDS), 0.1mol / L sodium chloride (NaCl), 10mmol / L triisopropyl ethylsulfonyl hydrochloride buffer (Tris-HCl, pH 8.0), 1 mmol / L ethylenediaminetetraacetic acid (EDTA, pH 8.0)) for 30 min, 10 μl of 10% proteinase K, and overnight in a water bath at 55°C.
[0028] (2) Add an equal volume of Tris saturated phenol (pH8.0) to the digested meat sample, slowly invert the centrifuge tube for 10 minutes, centrifuge at 12000rpm for 10 minutes, carefully draw the supernatant into a clean centrifuge tube with a large-caliber pipette tip .
[0029] (3) Repeat the previous step once. If the supernatant is clear and transparent, this step can be omitted.
[0030] (4) Add an equal amount of phenol:chloroform (1:1), slowly invert the centrifuge tube for 10 min, centrifuge at 12000 rpm for 10 min, and carefully draw ...
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
Login to View More Abstract
The invention discloses a method for rapidly detecting a quail-derived ingredient in food and a special primer. The special primer is shown in SEQ ID NO 1-2 in a sequence table. Quail meat-derived DNA is taken as a positive template, DNA of eight species, namely cattle, sheep, pigs, rabbits, pigeons, chicken, ducks and gooses, are taken as a hybrid template of an interference template, and PCR and qPCR reactions are respectively carried out, wherein a result of a PCR method takes an electrophoretic band in an agarose gel electrophoresis map as a positive result, the qPCR method takes amplification with Ct value of an amplification curve less than 34 as the positive result, and detection sensitivity reaches pictogram-scale DNA. The method disclosed by the invention has the advantages that 16SrRNA is utilized as a target gene for the first time to detect deeply processed quail meat, and the designed primer is applicable to PCR and qPCR methods at the same time, so that the method disclosed by the invention has good specificity, high sensitivity explores a new approach for identification of an animal derived ingredient in the food, and the method disclosed by the invention has the beneficial effects of convenience in operation, stability and accuracy.
Description
technical field [0001] The invention belongs to the technical field of food quality and safety detection, in particular to quail-specific PCR amplification identification of quail meat-derived components in livestock and poultry meat. Background technique [0002] In recent years, various food safety incidents have occurred frequently, such as adulteration of animal-derived food, shoddy quality, and the substitution of cheap and low-quality livestock and poultry meat for high-priced high-quality livestock and poultry meat, which seriously disrupted the market order and harmed producers and consumers. The legitimate interests of consumers have also brought serious food safety risks. In the West, a test and analysis of nearly a thousand kinds of meat products showed that nearly 20% of the products had labels that did not completely match the varieties. [0003] The biggest characteristic of quail belonging to Galliformes is resistance to rough feeding, strong disease resistan...
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
Login to View More Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2565/125
Inventor 樊艳凤高玉时唐修君葛庆联顾荣贾晓旭刘茵茵马丽娜张静黄胜海王珏
Owner JIANGSU INST OF POULTRY SCI