Characteristic sequence, primers and method for identifying Polygonatum filipe and Polygonatum cyrtonema
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A technology of Polygonatum polyflora and characteristic sequences, which can be applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., and can solve the problems of multiple PCR amplification map bands, poor repeatability, and low specificity. , to achieve the effect of a simple method
Active Publication Date: 2019-03-15
ZHEJIANG FORESTRY ACAD +1
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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.
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Embodiment 1
[0030] (1) extraction of genomic DNA of two kinds of Polygonatum:
[0031] 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.
[0032] (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.
[0033] (3) Design specific PCR amplifi...
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Abstract
The invention relates to a characteristic sequence and specific marker primers for identifying Polygonatum filipe and Polygonatum cyrtonema as well as a rapid identification method. The characteristicsequence is shown as SEQ ID NO.1; the primer sequences are shown as follows: the sequence of an upstream primer is 826F1:5'-ACACACACCACTAATGAAA-3', and the sequence of a downstream primer is 826R1:5'-AAACTACTAAGAGAGCGTGAG-3'. The characteristic sequence and molecular specific marker primers disclosed by the invention can be used for rapidly identifying the Polygonatum filipe and the Polygonatum cyrtonema. The method is simple, fast and accurate, and is an irreplaceable molecular means for distinguishing the Polygonatum filipe and the Polygonatum cyrtonema according to morphologic features.
Description
(1) Technical field [0001] The invention relates to a characteristic sequence for distinguishing Polygonatum filipe and Polygonatum cyrtonema, a specific marker primer, and a method for accurately distinguishing the two by using the primer. (2) Background technology [0002] 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 medicinal and edi...
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