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Characteristic sequence, primers and method for distinguishing Polygonatum long-stemmed and Polygonatum polyflora

A Polygonatum Polygonatum and Rhizoma Polygonatum technology, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of many bands, poor repeatability, and low specificity in PCR amplification patterns, etc. Achieving simple results

Active Publication Date: 2021-05-14
ZHEJIANG FORESTRY ACAD +1
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AI Technical Summary

Problems solved by technology

These molecular marker technologies all use general-purpose primers, which require a large number of primers to screen, and there are many bands in the PCR amplification pattern, with poor reproducibility and low specificity.

Method used

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  • Characteristic sequence, primers and method for distinguishing Polygonatum long-stemmed and Polygonatum polyflora
  • Characteristic sequence, primers and method for distinguishing Polygonatum long-stemmed and Polygonatum polyflora
  • Characteristic sequence, primers and method for distinguishing Polygonatum long-stemmed and Polygonatum polyflora

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] (1) extraction of genomic DNA of two kinds of Polygonatum:

[0039] Take the tubers of Polygonatum to be tested, wash them, slice them, dry them on silica gel (≥7d), take 0.03g, add liquid nitrogen and grind them thoroughly, and the genomic DNA is extracted using a new rapid plant genomic DNA kit (DP3112, BioTeke, Beijing BioTek Biology Co., Ltd. Technology Co., Ltd.), according to the instructions to extract the genomic DNA of the obtained specimens.

[0040] (2) The integrity, purity and concentration of the obtained genomic DNA were detected by 1.5% agarose gel electrophoresis and a DNA / RNA ultraviolet spectrophotometer (Nanodrop Technologies, USA). OD 260 / OD 280 DNA samples >1.8 were used for subsequent PCR amplification, and were appropriately diluted according to the nucleic acid concentration value, so that the genomic DNA concentration was about 20ng / μL. DNA extracts were stored in a -20°C refrigerator for later use.

[0041](3) Design specific PCR amplific...

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Abstract

The invention relates to a characteristic sequence for distinguishing Polygonatum filipe and Polygonatum cyrtonema, a specific marker primer and a rapid identification method. The characteristic sequence is shown in SEQ ID NO.1, and the primer sequences are as follows: upstream primer 891F2: 5′-TTCGGTTGTAGTGGTGCTTG-3′, downstream primer 891R2: 5′-AGGCTTGTACAATAAATCTAGAAAC-3′. The characteristic sequence and molecular specific marker primers of the invention can quickly identify the Polygonatum long stem and Polygonatum polyflora, the method is simple, fast and accurate, and is an irreplaceable molecular means for distinguishing Polygonatum long stem and Polygonatum polyflora by appearance characteristics.

Description

[0001] (1) Technical field [0002] The invention relates to a characteristic sequence for identifying Polygonatum filipe and Polygonatum cyrtonema, a specific marker primer, and a method for accurately distinguishing the two by using the primer. [0003] (2) Background technology [0004] Polygonatum, first recorded in "Bielu of Famous Doctors", is sweet in nature and flavor, flat, and returns to the spleen, lung, and kidney channels. Clinically, it has a certain effect on the treatment of fatigue, spleen and stomach qi deficiency, kidney deficiency, lung deficiency and dry cough, internal heat and thirst, deficiency of essence and blood, premature graying of beard and hair, dry mouth and lack of food, etc. It can also be used to treat tuberculosis, diabetes, chronic hepatitis and other diseases, and has anti-tumor and anti-radiation effects. Polygonatum underground tubers are rich in polysaccharides, saponins, amino acids, alkaloids, trace elements, etc., and have high medic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 叶碧欢李海波胡传久宋其岩陈友吾杜国坚余水生
Owner ZHEJIANG FORESTRY ACAD
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