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Specific primers for identification of Rhodiola crenulata at Mila mountain region in Tibet and identification method thereof

A technology of Rhodiola grandiflora and specific primers, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problem of no discovery and achieve short time consumption, low technical threshold, high The effect of accuracy

Active Publication Date: 2018-02-02
津宁植化(广州)科技发展有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The chemical composition of Rhodiola grandiflora in different regions of Tibet is very different. At present, most of the research on Rhodiola grandiflora in different regions of Tibet is based on the analysis of differences in certain chemical components and the examination of genetic diversity. After reading a large number of literature, There is no report on the detection of Rhodiola daflora in the Mila Mountain area of ​​Tibet

Method used

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  • Specific primers for identification of Rhodiola crenulata at Mila mountain region in Tibet and identification method thereof
  • Specific primers for identification of Rhodiola crenulata at Mila mountain region in Tibet and identification method thereof
  • Specific primers for identification of Rhodiola crenulata at Mila mountain region in Tibet and identification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Using the primer sequence:

[0025] 5'TCCGTTGCTGAATGTCTGTC3' and 5'ATCAAGCATGATTGTTGCGT3', the primer sequence is: SEQ ID No1-2. The specific steps for the detection method of Rhodiola grandiflora plants in the Mila Mountain area of ​​Tibet are as follows:

[0026] (1) Use the improved CTAB method to extract Rhodiola genomic DNA: Take 200 mg of young cauliflower leaves, grind them into powder in liquid nitrogen, transfer them to a 1.5 ml centrifuge tube, add 600 microliters of CTAB lysate, and incubate at 65°C for 30 Minutes, add 450 microliters of chloroform and 18 microliters of isoamyl alcohol, mix thoroughly, centrifuge, take the supernatant and add 1 times the volume of chloroform, mix well and centrifuge, take the supernatant and add 2 times the volume of ethanol, mix After uniformity, place it at -20°C for 30 minutes, centrifuge at 13000rmp for 15 minutes, remove the supernatant, rinse the precipitate with 75% ethanol and air-dry, add 200 microliters of 0.3mol / L...

Embodiment 2

[0032] Comparative Test

[0033]

[0034] The coincidence rate between the results identified by the above method and the planting area was 98%. The coincidence rate between the identification results of the conventional method and the planting area is 92%, which shows that the method of the present invention has higher accuracy, and the method has important application value in the identification of Rhodiola grandiflora in the Mila Mountain area of ​​Tibet.

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Abstract

The invention discloses sequences of a pair of specific primers for the identification of Rhodiola crenulata at Mila mountain region in Tibet and an identification method thereof, wherein, the sequences of the primers consist of 5'TCCGTTGCTGAATGTCTGTC3' and 5'ATCAAGCATGATTGTTGCGT3'.The method for the identification of Rhodiola crenulata at Mila mountain region in Tibet using the primers includes 4steps: (1) extraction of DNA from genome of five species of Rhodiola crenulata at different altitude in Tibet, (2) PCR amplification of specific primer pairs, (3) gel electrophoresis detection of PCRproducts; (4) determination of results: according to the location of a standard DNA marker belt on the gel, identifying whether the tested product is Rhodiola crenulataat Mila mountain region in Tibet or not. The coincidence rate of the results obtained by using the above-mentioned method to the planting area is more than 95%. The method has important application value in the identification of Rhodiola crenulata at Mila mountain region in Tibet.

Description

technical field [0001] The invention relates to a primer sequence for identifying Rhodiola grandiflora in the Mila mountain region of Tibet in biotechnology and a method for identifying Rhodiola grandiflora in the Mila mountain region using the primer sequence. Background technique [0002] Rhodiola big flower ( Rhodiola crenu Lata ) is Angiospermae (Angiospermae), Dicotyledoneae (Dicotyledoneae), Rosales (Rosales), CrassuLaceae (CrassuLaceae) Rhodiola ( Rhodiola L.) Plants. Rhodiola is a perennial herb that generally grows in mountainous areas with high altitudes of 3500-5000m, low temperature, and strong ultraviolet light. Many pharmacological studies at home and abroad have shown that Rhodiola has anti-aging, anti-fatigue, anti-tumor, and anti-oxidative properties. It has unique effects in sleep, sedation and physiological regulation, and is widely used in health care products, medicines, cosmetics and other fields. [0003] There are about 90 species of Rhodiola in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 刘志伟
Owner 津宁植化(广州)科技发展有限责任公司
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