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Sea cucumber peptide identification and detection kit and detection method thereof

A technology for detection kits and detection methods, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc. It can solve the problems of low molecular weight of peptides and the inability to determine the source of peptides, etc., and achieve simple and fast operation, simple identification and detection. Effect

Inactive Publication Date: 2018-05-01
DALIAN SHENLAN PEPTIDE TECH R & D CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the low molecular weight of the peptide, the source of the peptide cannot be determined by protein sequencing and alignment

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A sea cucumber peptide identification and detection kit, comprising the following components: a mixture of phenol and chloroform; absolute ethanol; specific primers; a PCR reaction mixture; trichloroacetic acid; a mixture of copper sulfate and potassium sulfate;

[0034] The sequence of the specific primer is:

[0035] Upstream primer: 5'- ACTAATACCTCCCCTCCTT -3'

[0036] Downstream primer: 5'-ATAAACTTCTGGGTGTCC-3'.

[0037] The volume ratio of phenol to chloroform in the phenol and chloroform mixture is 1:1.

[0038] The trichloroacetic acid is 9%-15% trichloroacetic acid.

[0039] The mass ratio of copper sulfate and potassium sulfate in the mixture of copper sulfate and potassium sulfate is 1:30.

[0040] The PCR reaction mixture is: containing Mg 2+ Mix 10×PCR buffer, dNTP, and 5U / μL TaqE at a volume ratio of 5:4:0.5.

[0041] The kit is used for: identifying sea cucumber peptides by PCR method and identifying and distinguishing sea cucumber peptides from other...

Embodiment 2

[0043] A detection method utilizing the sea cucumber peptide identification and detection kit described in Example 1: specifically comprising the following steps:

[0044] (1) Identification of sea cucumber peptides

[0045] a. Extraction of DNA from sea cucumber peptide samples:

[0046] Take 0.1g of sea cucumber peptide sample in a 1.5mL EP tube, add 600μL of phenol and chloroform mixture, gently invert for 10min, 12000rpm, centrifuge for 10min; take 500μL of supernatant, repeat the previous step twice, take supernatant 400μL, add 800μL absolute ethanol and shake well to precipitate DNA; 12000rpm, centrifuge for 10min;

[0047] b. PCR amplification of the COI gene fragment in sea cucumber peptide sample DNA:

[0048] Use the DNA in the extracted sea cucumber peptide sample as a template to amplify the COⅠ fragment by PCR. The 50 μL PCR reaction system is: 2 μL of DNA template, 2 μL of upstream and downstream primers, 9.5 μL of PCR reaction mixture, and 34.5 μL of water; ...

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PUM

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Abstract

The invention belongs to the field of food detection, and in particular relates to a sea cucumber peptide identification and detection kit and a detection method thereof. The sequences of a specific primer are: 5'-ACTAATACCTCCCTCCTT-3' (upstream) and 5'-ATAAACTTCTGGGTGTCC-3' (downstream). The sea cucumber peptide identification and detection kit is simple and rapid in operation by utilizing a PCRmethod, can rapidly identify sea cucumber peptide and can identify and differentiate the sea cucumber peptide and other peptide products; and the kit contains a peptide content detection reagent bottle which can measure the peptide content of the sea cucumber sample. Therefore, the sea cucumber peptide identification and detection kit can identify and detect the sea cucumber products rapidly, simply and conveniently.

Description

technical field [0001] The invention belongs to the field of food detection, and in particular relates to a sea cucumber peptide identification and detection kit and a detection method thereof. Background technique [0002] There are more than 1,400 species of sea cucumbers in the world, which are distributed in the vast sea areas of the intertidal zone and the ocean floor, among which the tropical sea area is rich in species. "Food Materia Medica" records that sea cucumber tastes salty, warm and tonic, enters the kidney meridian, generates blood in all veins, nourishes kidney and essence, strengthens yang and treats impotence, relieves labor and eliminates phlegm, nourishes yin and diuresis, corrects softness and firmness, clears intestines and moisturizes dryness. "Wuzazu" records that sea cucumbers are found on the coast of Liaodong, and their warm nature complements enemy ginseng, so it is called sea cucumbers. The protein content of dried sea cucumber is as high as mor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6876C12Q1/686
CPCC12Q1/6876C12Q1/686C12Q2565/125
Inventor 包卫洋马普王祖哲杨大苹
Owner DALIAN SHENLAN PEPTIDE TECH R & D CO LTD