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Method for conducting real-time fluorescence identification on edible bird's nest product through PCR

A real-time fluorescence and identification method technology, which is applied in the identification field of bird's nest, can solve the problems of low sensitivity, low accuracy, lengthy extraction and purification procedures, etc., and achieve the effect of good specificity and high sensitivity

Inactive Publication Date: 2016-05-18
英格尔检测技术服务(上海)有限公司 +1
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Problems solved by technology

Some of these methods have low accuracy and low sensitivity; some extraction and purification procedures are lengthy and have poor reproducibility; some identification costs are high

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  • Method for conducting real-time fluorescence identification on edible bird's nest product through PCR
  • Method for conducting real-time fluorescence identification on edible bird's nest product through PCR
  • Method for conducting real-time fluorescence identification on edible bird's nest product through PCR

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Embodiment Construction

[0057] In order to make the technical means, creative features, goals and effects achieved by the present invention easy to understand, the present invention will be further described below in conjunction with specific diagrams.

[0058] see figure 1 , figure 2 , image 3 , Figure 4 , Figure 5 A real-time fluorescent PCR identification method for bird's nest products, comprising the following steps: Step 1, DNA extraction of bird's nest products to be inspected; Step 2, real-time fluorescent PCR amplification of swiftlet components; first, adding 2 × PCRMasterMix12 to the PCR reaction tube .5 μL, 0.5-1 μL of upstream primer and downstream primer, 0.5-1 μL of Taqman probe, 1-2 μL of 50 ng / μL DNA of bird’s nest product to be tested, supplemented with double distilled water to 25 μL, and mixed well; secondly, put the PCR reaction tube into the fluorescent Quantitative PCR instrument, complete PCR amplification according to the following reaction conditions: a.94-95°C 5min,...

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Abstract

The invention relates to the technical field of chemical detection and discloses a method for conducting real-time fluorescence identification on an edible bird's nest product through PCR. The method comprises the following steps: 1, extracting DNA of the edible bird's nest product to be detected; 2, conducting real-time fluorescence PCR amplification on esculent swift constituents, wherein firstly, 12.5 microliters of 2*PCR Master Mix, 0.5-1 microliter of an upstream primer, 0.5-1 microliter of a downstream primer, 0.5-1 microliter of Taqman probes and 1-2 microliters of the edible bird's nest product to be detected with the concentration being 50 ng / microliter are added into a PCR reaction tube, supplementing double distilled water to be 25 microliters, and conducting uniform mixing; secondly, putting the PCR reaction tube into a fluorescent quantitative PCR instrument to complete PCR amplification; 3, analyzing the amplification result with real-time fluorescence PCR instrument analysis software. The main principle of the method is that an edible bird's nest is mainly from saliva of esculent swifts and contains DNA of the esculent swifts, and realness of the edible bird's nest is identified by detecting the 12S rRNA gene of the esculent swifts in the edible bird's nest.

Description

technical field [0001] The invention relates to the technical field of chemical detection, in particular to an identification method of bird's nest. Background technique [0002] Bird's nest is one of the traditional and precious foods that have been eaten in China since the Ming Dynasty. It refers to the saliva secreted by the insects, seaweed, whitebait, etc. eaten by several swiftlets of the swiftlet family Swiftletaceae. Nests mixed with down feathers and the like. The swallow's nest is half-moon-shaped, with a diameter of 6-7 cm, a thick base, a thin wall, and a weight of about 5-15 grams. The bird's nest is neat on the outside and rough on the inside, like a network of loofahs. It is mainly produced in Southeast Asian countries such as Indonesia, Malaysia, Thailand and Vietnam. [0003] Traditional Chinese medicine believes that bird's nest: "nourishing yin and moistening dryness, nourishing qi and nourishing the middle, treating deficiency, coughing and asthma, hem...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2561/101C12Q2561/113
Inventor 赵玉簪
Owner 英格尔检测技术服务(上海)有限公司
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