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Seahorse identification method

An identification method and hippocampal technology, applied in the field of molecular biology, can solve the problems of cumbersome, restricting the use and promotion of molecular identification methods, and time-consuming, and achieve the effects of protecting health, clear test results, and simple operation.

Inactive Publication Date: 2015-09-09
CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For a long time, DNA extraction has been the most time-consuming and cumbersome step in DNA analysis technology experiments, which seriously restricts the use and promotion of molecular identification methods

Method used

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  • Seahorse identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] 1. DNA extraction method from hippocampus

[0090] 1.1 Materials and methods

[0091] 1.1.1 Materials

[0092] The materials used in the experiment are shown in Table 1, including:

[0093] Table 1 Basic information of hippocampus-related samples

[0094]

[0095] Note: "*" means hippocampus recorded in Pharmacopoeia, this study uses the name contained in "Chinese Fish Systematic Retrieval", and the name in brackets is Pharmacopoeia name.

[0096] 1.1.2 Experimental Instruments

[0097] Icycle PCR instrument (Bio-Rad, USA); centrifuge, constant temperature metal bath, ultraviolet lamp

[0098] 1.1.3 Method

[0099] Template DNA extraction: Select 50 mg of dry medicinal material specimens without mildew, cut them into powder with scissors, transfer them to a 1.5 mL microcentrifuge tube, add 60 μL of alkaline lysis buffer (0.5 mol / L NaOH, 1% PVP-40, 1% Triton X-100); use a vortex shaker to mix for 20s, boil for 30s, take out; add 0.1M Tris-HCl (pH=8) 200uL, vortex ...

Embodiment 2

[0105] Template DNA was prepared by the method in Example 1.

[0106] The total PCR amplification reaction system is 25 μL, including 12.5 μL of 2x Taq master mix, 0.5 μL of each primer (Mo-F / Mo-R, Ku-F / Ku-R, Tr-F / Tr-R) (20 μmol / L ), template 1μL (about 30ng), add ddH2O to make up to 25μL. Pre-denaturation at 94°C for 1min; denaturation at 94°C for 30s, 30s at 64°C, a total of 30 cycles. After the reaction was completed, 1 μL of 100-fold diluted SYBR Green I dye was added, and the color of the reaction product of the tested sample was detected under ultraviolet light. Another 5 μL product was taken, and the bands were detected by 1.5% agarose gel electrophoresis, and observed by ultraviolet gel imaging, such as Figure 4 .

[0107] 2. Results

[0108] Under ultraviolet fluorescence inspection, if the color is green or yellow-green, it means that at least one of Japanese hippocampus, tube hippocampus, and three-spotted hippocampus exists in the tested medicinal materials, a...

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Abstract

The invention relates to a seahorse identification method, and belongs to the field of molecular biology. The technical problem to be solved is to provide a seahorse identification method. The seahorse identification method can realize the accurate identification of hippocampus japonicus, hippocampus trimaculatus and hippocampus kuda. The seahorse identification method comprises the following steps of: a. extracting seahorse DNA (deoxyribonucleic acid); b. carrying out PCR (Polymerase Chain Reaction) amplification on the seahorse DNA; and c. after ending the PCR amplification, adding 1 microlitre of 100X SYBR Green I dye to a reaction system, and detecting the color of the sample under an ultraviolet lamp; if the sample is not yellow or yellow green, the sample does not contain the corresponding seahorse, ending identification; and if the sample is yellow or yellow green, the sample contains the corresponding seahorse, taking 5 microlitre of PCR product to carry out 1.5% agarose gel electrophoresis detection, and identifying the seahorse variety.

Description

technical field [0001] The invention relates to a method for identifying hippocampus and belongs to the field of molecular biology. Background technique [0002] Seahorse is a general term for fishes of Osteichthyes, Syngnathiformes, Syngnathidae, and Hippocampus. It is mainly used as traditional medicinal materials in East Asia. Hippocampus is a traditional and precious Chinese medicinal material in my country, which was first recorded in "The Collection of Notes on Materia Medica". The 2010 edition of "Chinese Pharmacopoeia" recorded hippocampus as Hippocampus. "Chinese Fish System Retrieval" records that H.kelloggi is a large hippocampus, H.kuda is a tube hippocampus, and H.japonicus is a Japanese hippocampus H.mohnkei. This study uses "Chinese Fish Retrieval". Seahorse name) are included in CITES (the Convention on Internaitional Trade in Endangered Species of Wild Fauna and Flora) Appendix II. [0003] There are about 33 species of seahorses in the world, and 35 spec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q1/686C12Q2537/143C12Q2563/107C12Q2527/101
Inventor 国锦琳袁媛王茜侯飞侠彭成
Owner CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE
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