Banding detection method of Cordyceps sinensis DNA fingerprint high resolution ratio
A DNA fingerprinting and detection method technology, applied in the field of molecular biology, can solve the problems of large amount of Cordyceps sinensis identification and identification methods, insufficient accuracy and high efficiency, etc., and achieves the effects of a reliable and efficient evaluation system, shortening time, and improving resolution.
Active Publication Date: 2010-10-06
NAVY MEDICINE RES INST OF PLA
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Problems solved by technology
[0004] The technical problem to be solved by the present invention is to solve the existing Cordyceps sinensis identification and identification method with a large amount of use, not accurate and efficient, and many defects in the radioactive banding method and the fluorescent banding method, and to establish a high-resolution display that can fully adapt to the high-resolution DNA fingerprint of Cordyceps sinensis. with detection method
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Embodiment 1
[0013] (1) PAGE gel electrophoresis step of Cordyceps sinensis DNA: use 3%-6% PAGE gel to detect the DNA fingerprint of Cordyceps sinensis.
[0014] Table 1 Concentration table of polyacrylamide gel
[0015]
[0016] Prepare polyacrylamide gels with different concentrations according to the above table, extract Cordyceps sinensis DNA according to conventional methods, and perform PAGE gel electrophoresis.
[0017] (2) Glue fixation process: its purpose is to make nucleic acid better and Ag + For specific binding, the fixation time is 15-30min, and instead of 10% ethanol, 0.5% acetic acid is directly used in the fixative concentration, which obviously improves the specificity.
[0018] (3) The staining solution uses 0.1%-0.4% AgNO 3 Solution formulation. Remove and separate the PAGE gel plate, put the side of the PAGE gel with the DNA fingerprint of Cordyceps sinensis into ddH 2 Rinse in 0 (2000ml) for 3-6s, then put the PAGE plate into a container filled with staining s...
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The invention belongs to the technical field of the molecule biology, and particularly relates to a banding detection method for valuable endangered medical plant Cordyceps sinensis DAN fingerprint high resolution. The invention discloses a banding detection method of Cordyceps sinensis DNA fingerprint high resolution ratio, which comprises a PAGE gel electrophoresis step, a glue fixing step, a glue dying step and a glue color development step of Cordyceps sinensis DAN, and is characterized in that fixing liquid in the glue fixing step is of 0.5 percent acetic acid, dying liquid in the glue dying step is of 0.1 to 0.3 percent AgNO3, and developer solution in the glue color development step is of 1 to 4 percent strong base and 0.5 percent formaldehyde solution. The banding detection method can simplify the steps and can save the time, can save the experiment reagent and material in the experiment, can obtain high-efficient and accurate Cordyceps sinensis high resolution PAGE banding so as to set a molecule analysis foundation for establishing the valuable endangered traditional Chinese medicine - cordyceps sinensis resource sustainable development system and to provide reliable and high-efficient evaluation system for the Cordyceps sinensis character index - chemistry - molecule system.
Description
technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a high-resolution band detection method for the DNA fingerprint of the rare and endangered medicinal plant Cordyceps sinensis. Background technique [0002] Cordyceps Cordyceps (Fr.) Link is a large class of entomopathogenic fungi, and Cordyceps sinensis (Berk.) Sacc.], which has important medicinal value after its specific fungus infects specific insects, is a precious medicinal material in my country. Cordyceps sinensis is sweet and flat in nature and taste, enters the lung and kidney meridian, can benefit lung and kidney, relieve cough, tonify deficiency and damage, benefit essence, anti-aging, anti-cancer, and has the effect of enhancing human immune function and regulating reproduction. Natural Cordyceps sinensis is mainly produced in Tibet, Yunnan, Sichuan and Qinghai-Tibet, with rich germplasm resources. However, there are many kinds of fakes of the ...
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IPC IPC(8): A61K36/068C12Q1/68G01N1/30G01N27/447A61P1/14A61P11/14A61P35/00A61P37/04A61P39/06
Inventor 张阵阵汪家春储智勇谷爱梅张淼
Owner NAVY MEDICINE RES INST OF PLA