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Extraction method of rheum tanguticum

An extraction method and technology for medicinal materials, which are applied in the field of DNA extraction of Tanggut rhubarb medicinal materials, can solve the problems of high cost, low DNA purity, high price and the like, and achieve the effects of low cost, simple process and convenient operation.

Inactive Publication Date: 2010-12-22
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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Problems solved by technology

However, no matter which method is used to pretreat the medicinal materials, there are DNA fragments in the electrophoresis detection of the extracted DNA, but the extracted DNA has tailing phenomenon, indicating that the degradation is serious
Gained DNA is carried out purity test with Beckman coulter DU640 ultraviolet spectrophotometer in addition, the ratio (A) of absorbance (A) at 260nm and 280nm 260 / A 280 ) can be used as an indicator of DNA purity, and the A 260 / A 280 Between 1.65-1.83; soak the medicinal material treated with 0 ℃ extraction medium I, the A of the obtained DNA extract 260 / A 280 Between 1.38-1.57, indicating that the resulting DNA purity is low
[0005] Komatsu Katsuko (2006) used the DNeasyTM Plant Mini Kit (QIAGEN, Germany) kit to extract the genomic DNA from rhubarb medicinal materials from different sources, and the kit is relatively expensive and expensive

Method used

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  • Extraction method of rheum tanguticum
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Embodiment Construction

[0029] 1. Extraction of DNA from rhubarb tangutica

[0030] (1) Collect Tangut rhubarb root in the wild, cut it into sections every 3 cm, thread it with a thread and dry it naturally for later use.

[0031] Take 2g of Tangut rhubarb medicinal material and soak it in 300ml of 0℃ distilled water, change the distilled water every 4h, soak for 24h; then cut into small pieces, dry in air for later use. Wipe the surface with 70% ethanol or gently scrape off the surface contamination before use.

[0032] (2) Take 0.1 g of the pretreated rhubarb Tanguta medicinal material and grind it into powder in a mortar with a little quartz sand, and add a little PVP dry powder while grinding.

[0033] (3) Transfer the ground dry powder into a 2ml centrifuge tube, add 1ml of ice-cold extraction medium I (mixture of 200mmol / L Tris-HCl, pH8.0, 50mmol / L EDTA pH8.0, 0.25mol / L NaCl) and 10 μl β-mercaptoethanol at 0 ° C for 10 min. Centrifuge at 2000r / min for 10min at 4°C, collect the precipitate, t...

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Abstract

The present invention provides a method for extracting a DNA from Rheum tanguticum Maxim. The method includes the following steps: after being soaked in distilled water for 24 hours, the dry Rheum tanguticum Maxim is cut into small pieces, dried and then ground into powder; the medicine powder is extracted by extraction medium I and Beta-mercaptoethanol and then by extraction medium II and Beta-mercaptoethanol; after the product is extracted by chloroform / isoamylol, the aqueous phase is added into NaAc(pH5.2) and ice-cold isopropanol in order to precipitate DNA; the aqueous phase is discarded; the precipitate is washed by 75 percent of ethanol for two to three times and then naturally aired after the ethanol is removed; the DNA is dissolved in sterilizing ddH2O; and the solution is kept under the temperature of 4 DEG C or placed under the temperature of 20 DEG C below zero for a long time. Being tested by electrophoresis, the extracted Rheum tanguticum Maxim. DNA is an intact genome DNA, and the purity is high (A260 / 280 equal to 1.8 to 20). Serving as a template, the obtained medicine genome DNA is amplified respectively by UBC888 primer and UBC891 primer, so that a map with clearstrips can be obtained.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a method for extracting DNA from medicinal materials, in particular to a method for extracting DNA from rhubarb tangutiae medicinal materials. Background technique [0002] Tangut rhubarb, also known as rhubarb with chicken feet, belongs to the genus RheumL of the Polygonaceae family. It is a perennial herb distributed in eastern Tibet, Qinghai, and Gansu. In the middle and semi-shady slopes, it is a unique species in China. Its dried roots and rhizomes are used as medicine, cold in nature and bitter in taste; it has the effects of purging heat and dredging intestines, cooling blood and detoxifying, removing blood stasis and stimulating menstruation; it is one of the "authentic rhubarb" recorded in "Pharmacopoeia of the People's Republic of China" (2005). [0003] With the development of molecular biology, DNA molecular marker technology has gradually penetrated into the field...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10C07H1/06
Inventor 李毅胡延萍王莉杨建谢小龙
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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