Primers, probes and kits for simultaneous detection of goat and cow origin in raw milk
A kit and goat technology, applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problem of less multi-channel and multi-source detection, and achieve the effect of reducing mismatches
Active Publication Date: 2020-08-04
XILINGOL VOCATIONAL COLLEGE
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Problems solved by technology
At present, the technical research, testing standards, invention patents and commercial kits related to the authenticity of dairy products are mainly focused on single-channel single-source detection and heterogeneous quality control detection, while multi-channel multi-source detection and same-tube quality control detection Less reported
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[0063] 1. Detection method:
[0064] (1) Extract the DNA of raw milk, and set up an extraction blank control (the control group for subsequent extraction methods).
[0065] (2) Detect the concentration and quality of DNA, and dilute the concentration to 100-200ng / μL.
[0066] (3) Utilize multiple fluorescent quantitative PCR primers and probes to carry out amplification detection on the DNA dilution, use goat and cow positive standards as positive controls, use sterilized deionized water as negative controls, and use DNA extraction blank controls as For the control group of the extraction method, the Real-time PCR reaction system is shown in Table 1, and the Real-time PCR amplification parameters are shown in Table 4.
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The invention discloses primers and probes for synchronously detecting goat origin and cow origin in raw milk as well as a kit. The sequences of the primers and the probes are as follows: the sequenceof a goat origin detection forward primer is as shown as SEQ ID No.1; the sequence of a cow origin detection forward primer is as shown as SEQ ID No.2; the sequence of a goat origin detection reverseprimer is as shown in SEQ ID No.3; the sequence of a cow origin detection reverse primer is as shown in SEQ ID No.4; the sequence of a goat probe is as shown in SEQ ID No.5; the sequence of a cow probe is as shown in SEQ ID No.6; and the sequence of a quality control probe is as shown in SEQ ID No.7. The primers, the probes and the kit have high specificity and high sensitivity, can realize same-tube detection on goat origin and cow origin in the raw milk and quality control, and can perform quantitative detection on goat origin and cow origin.
Description
technical field [0001] The invention belongs to the technical field of food detection, in particular to the field of detection of animal origin in raw milk. Background technique [0002] Dairy products are an important part of human dietary structure, rich in nutrients such as protein, fat, carbohydrates, vitamins and minerals (including trace elements). These nutrients are not only the basis of macroscopic human body and microscopic cells, but also the source of regulating human life activities and intracellular biochemical reactions. In addition to being rich in conventional nutrients, dairy products have a sensory experience of good color, aroma and taste, which endows them with the core of global cuisine. However, with the increasing production and consumption, dairy products have become the main target of fraud and adulteration in the fields of food production, processing, distribution and catering. [0003] In our country, the research on the authenticity identificat...
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IPC IPC(8): C12Q1/6888C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6888C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2563/107C12Q2545/114
Inventor 郭梁郭元晟巨艳鲁铁冯伟廖成松雅梅钱俊平孙建萍朱建军
Owner XILINGOL VOCATIONAL COLLEGE



