Chinese traditional medicine armand clematis polyase polymerase chain reactionidentification primer, reagent kit and identification method
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A kind of Chuanketong and polymerase technology, which is applied in the direction of material analysis, instruments, and analysis materials through electromagnetic means, can solve the problems of different identification methods, and achieve the effect of high reproducibility and strong stability
Inactive Publication Date: 2009-12-09
CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE
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Problems solved by technology
However, this method is not the same for different Chinese medicinal materials, the primers for identification and the method for identification
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Embodiment 1
[0045] The preparation of embodiment 1 kit of the present invention
[0046] a. Specific identification primers: upstream primer: 5′-TGCCCAGCCTGCACAAGA-3′, downstream primer: 5′-TGCCCAGCCTCAACAGTGT-3′, the primers were synthesized by an automatic DNA synthesizer; each 1.0 μl;
[0047] b. Positive control substance: 1.0 μl of Chuanketong DNA template;
[0048] c. 1 unit of TaqDNA polymerase, 10×PCR reaction buffer, MgCl 2 , dNTP;
[0049] D, DNA extraction reagent; (the reagent that the present invention adopts all adopts commercially available routine reagent)
[0050] Among them, 10×PCR reaction buffer 2.5μl, 25mM MgCl 2 2.0 μl, 2.5 mM dNTP 1.5 μl.
Embodiment 2
[0051] Embodiment 2 Preparation of kit of the present invention
[0052] a. Specific identification primers: upstream primer: 5′-TGCCCCAGCCTGCACAAGA-3′, downstream primer: 5′-TGCCCAGCCTCAACAGTGT-3′, the primers were synthesized by an automatic DNA synthesizer; each 0.5 μl;
[0053] b. Positive control substance: 1.0 μl of Chuanketong DNA template;
[0054] c. 1 unit of TaqDNA polymerase, 10×PCR reaction buffer, MgCl 2 , dNTP (commercially available);
[0055] d, DNA extraction reagent (commercially available);
[0056] Among them, 10×PCR reaction buffer 2μl, 25mM MgCl 2 1.5 μl, 2.5 mM dNTP 1.0 μl.
Embodiment 3
[0057] Embodiment 3 Preparation of kit of the present invention
[0058]a. Specific identification primers: upstream primer: 5′-TGCCCAGCCTGCACAAGA-3′, downstream primer: 5′-TGCCCAGCCTCAACAGTGT-3′, the primers were synthesized by an automatic DNA synthesizer; each 1.5 μl;
[0059] b. Positive control substance: 2.0 μl of Chuanketong DNA template;
[0060] c. 1 unit of TaqDNA polymerase, 10×PCR reaction buffer, MgCl 2 , dNTP (commercially available);
[0061] d, DNA extraction reagent (commercially available);
[0062] Among them, 10×PCR reaction buffer 3μl, 25mM MgCl 2 3.0 μl, 2.5 μl of 2.5 mM dNTP.
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Abstract
The invention relates to an identification primer for the chain reaction of the Chinese traditional medicine of armand clematis polymerase; the DNA sequence of the identification primer is that the sequence of an upstream primer is 5'-TGCCCAGCCTGCACAAGA-3' and the sequence of a downstream primer is 5'-TGCCCAGCCTCAACAGTGT-3'. The invention also provides a method for adopting the identification primer to identify the Chinese traditional medicine of the armand clematis polymerase and a reagent box which is used for identifying whether the armand clematis polymerase is authentic or not. The invention solves the problem that the quality goods of the armand clematis polymerase is hard for identification. A pair of high-special primers is designed for identifying the armand clematis polymerase in the process of identifying the armand clematis polymerase by the inventor; the identification primer can only specialally identify the armand clematis polymerase under preset PCR conditions and can not magnify the segments of adulterant of the armand clematis polymerase. When the sample is supplied and magnified with the identification primer in the pattern of PCR; the sample is checked by agarose electrophoresis whether the magnifying strip exist to accurately identify the authenticity of the medicine.
Description
technical field [0001] The invention relates to the identification of the traditional Chinese medicine Chuanketong, in particular, it uses polymerase chain reaction identification primers to identify the traditional Chinese medicinal material, and belongs to the field of medicine. Background technique [0002] Chuanmutong is a commonly used traditional Chinese medicine, which has the functions of clearing away heat and diuresis, promoting menstruation and breast milk, etc. It has a long history of medicinal use. It is recorded as Clematis. armandii Franch.) or the stem of C. Montana Buch-Ham. However, the stems and vines of more than 13 species of Ranunculaceae are used as common trees in different regions to varying degrees, and the use of medicinal materials is mixed, which is related to the safety and effectiveness of clinical medication. Since the Akebia medicinal materials in the market are all clematis plants of the family Ranunculaceae, the medicinal materials themse...
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Application Information
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N21/33G01N27/447
Inventor 万德光国锦琳
Owner CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE
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