LAMP primer set and detection method for duck-derived components
A detection method and primer set technology, applied in biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc., can solve the problems of difficult on-site detection, many experimental steps, long time required, etc. Short, highly sensitive, easy-to-use effects
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0067] Example 1 The establishment of the duck-derived component LAMP detection method comprises the following steps
[0068] In this example, the DNA of chicken, duck, beef, mutton, beef cubes and mutton rolls were extracted by phenol-chloroform extraction method (refer to the "Molecular Cloning Experiment Guide" for the extraction method) and the kit method, and the extracted DNA was stored. Store in a -20°C refrigerator, take out a part and dilute to 25ng / ul, and place it in a 4°C refrigerator for use as a template for PCR amplification to avoid DNA degradation caused by repeated freezing and thawing.
[0069] 1. Experimental materials
[0070] Domestic duck (the mallard of Anserinae Anatiae), mutton, beef, chicken, and pork are all purchased in local markets in Shanghai; frozen mutton rolls, meat skewers, beef cubes, beef jerky and other processed livestock products are purchased In supermarkets and farmer's markets in Shanghai.
[0071] 2. LAMP primer design
[0072] A...
Embodiment 2
[0107] Example 2 Primer specificity and sensitivity analysis of duck-derived component LAMP detection method
[0108] 1. LAMP primer specificity
[0109] The duck LAMP primer set designed in the present invention is used to carry out PCR amplification in mutton, beef, pork, chicken and different varieties of duck respectively, and carry out specificity experiments.
[0110] Extract the DNA of duck, beef, pork, chicken, and mutton, and use it as a template for the LAMP reaction. Perform LAMP-specific detection according to the optimized LAMP reaction conditions in Example 1. After the LAMP amplification reaction is completed, add SYBRGreen I to the amplification product Dye, observe the color change: the sample with green color is positive and contains duck-derived components; the sample with orange color is negative and does not contain duck-derived components, the result is as follows Figure 9 shown.
[0111] Depend on Figure 9 It can be seen that only duck meat amplifie...
Embodiment 3
[0119] Example 3 Sampling investigation of beef and mutton products in the market
[0120] Twenty-one samples were collected from 3 supermarkets and 2 farmers' markets in Minhang District, Shanghai, including five-spice stewed beef, brine beef, fatty beef, mutton rolls, mutton skewers, beef sauce, and beef cubes.
[0121] The LAMP detection method of the present invention is used to carry out sampling detection on the beef and mutton food in the market, and the verification is carried out by conventional PCR technology.
[0122] DNA was extracted from mutton rolls, beef rolls and beef cubes bought in the market, and then detected using PCR-specific primers and LAMP primers respectively. The experiments were carried out by two technicians at the same time, and the samples were tested according to the SNT1119-2002 standard. For the detection of cattle and sheep components, the detection results of conventional PCR technology and the LAMP detection method of the present invention...
PUM
Property | Measurement | Unit |
---|---|---|
Sensitivity | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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