Extraction method of rheum tanguticum

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 clear strips 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 and rock piles, 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...

AI Technical Summary

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 its genomic DNA from rhubarb medicinal materials from different sources, and the kit is relatively expensive and expensive

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