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A set of pcr identification primers and a method for identifying white and white chrysanthemums
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A technology of white and yellow flowers, applied in the fields of biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc.
Active Publication Date: 2017-02-22
贵州中医药大学 +1
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Before submitting the application of the present invention, there was no disclosure or report of the PCR identification primer set mentioned in this patent application and the molecular identification method for synchronous identification of white and yellow-flowered white and
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Embodiment 1
[0056] 1) Material
[0057] a. 27 samples of white and medicinal materials were collected in the medicinal material market, each sample was about 250g.
[0058] b. Reagent agarose (Biowestagarose, Spain); D2000DNA Marker (Beijing Suo Laibao Technology Co., Ltd.); 2 × Power Taq PCR MasterMix (Beijing Biotech Biotechnology Co., Ltd.); ethidium bromide (Beijing Tiangen Biochemical Technology Co., Ltd. ); primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd.
[0059] c. Instrument PCR instrument (Applied Biosystems Veriti TM 96, ABI, USA); gel imaging system (GGM / D2, SYNGENE, UK); computer Sanheng multipurpose electrophoresis instrument (DYY-12, Beijing Liuyi Instrument Factory).
[0060] 2) Genomic DNA extraction: 10 mg of the sample to be identified was taken, and the genomic DNA of each medicinal material sample was extracted by alkaline extraction, and stored at -20°C.
[0061] 3) Specific PCR amplification:
[0062] Use the specific primers shown in SEQ ID N...
Embodiment 2
[0071] 1) Materials, reagents and instruments
[0072] a.Material
[0073] 43 batches of plant samples of the same family or genus were collected, totaling 346 samples. Among them, there are 8 batches of Baiji samples, with a total of 72 samples; 10 batches of Huanghuabaiji, with a total of 103 samples; 2 species of the same plant (Xiaobaiji, Huabaiji), 8 batches, a total of 45 plants Samples: 17 batches of 4 species of plants of the same family (Solium chinensis, Orchid sylvestris, Orchid japonica, and Orchid basilicum), with a total of 126 samples. The number of samples and their sources are shown in Table 1.
[0074] Table 1 Sample source list
[0075]
[0076]
[0077] b. Reagent agarose (Biowestagarose, Spain); D2000 (Beijing Suo Lai Bao Technology Co., Ltd.); 2 × Power Taq PCR MasterMix (Beijing Biotech Biotechnology Co., Ltd.); ethidium bromide (Beijing Tiangen Biochemical Technology Co., Ltd.) ; Primers were synthesized by Shanghai Sangon Biotechnology Co., L...
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Abstract
The invention discloses a group of PCR identification primers. The group of primers comprises three primers, wherein the nucleotide sequences of the three primers are respectively shown in SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3 in a sequence table. The invention also provides a method for identifying B.striata Rchb.f. and B.ochracea Schltr. by using the primer group. The method comprises the following steps: 1) extracting genome DNA of samples to be identified; 2) by using the genome DNA extracted in the step 1) as a template, carrying out PCR amplification on the genome DNA by using the primer group; and 3) carrying out electrophoretic detection on the PCR amplification products obtained in the step 2), wherein the samples to be identified, which have stripes in 354bp, are B.striata Rchb.f., the samples to be identified, which have stripes in 519bp are as B.ochracea Schltr., and the samples to be identified, which have stripes in 354bp and 519bp are other plant materials. The method provided by the invention can be used for quickly and accurately identifying original plants, decoction piece medicinal materials and powdery medicinal materials of B.striata Rchb.f., synchronously identifying original plants, decoction piece medicinal materials and powdery medicinal materials of B.ochracea Schltr. and achieving identification of other plants.
Description
technical field [0001] The invention relates to the technical field of molecular markers, in particular to a set of PCR identification primers and a method for identifying white and white chrysanthemums. Background technique [0002] The traditional Chinese medicine Baiji has the effects of astringent and hemostasis, detumescence and granulation, and can be used for the treatment of hemoptysis, hematemesis, traumatic bleeding, sores and swelling, and chapped skin. All previous editions of the Chinese Pharmacopoeia have recorded Bletilla striata and Bletilla striata Rchb.f., which are rare and endangered plants, and the wild resources are scarce, and the cultivated resources are still difficult to meet the market demand, causing the price of medicinal materials to increase year by year. But because the distribution of this species has regional characteristics, some regional Chinese medicinal materials standards use Huanghuabai and B.ochracea Schltr. plants as the alternative ...
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