Quick extraction method for nucleic acid of diseased material of animal tissues
An extraction method and animal tissue technology, which are applied in the field of rapid extraction of nucleic acid from animal tissue disease materials, can solve the problems of long detection time, potential safety hazards due to the presence of toxic and harmful substances in extraction reagents, unsuitable for outdoor environment, etc. Low experimental equipment requirements, low cost effect
Inactive Publication Date: 2018-10-19
芜湖帮许来诺医疗设备科技有限公司
View PDF1 Cites 0 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
[0003] The purpose of the present invention is to provide a method for rapid extraction of nucleic acid from animal tissue disease materials. By improving the lysate used in the nucleic acid extraction process, the nucleic acid extraction method is greatly simplified. The operation is simple and safe, and the cost is low. The nucleic acid extraction method is not suitable for the outdoor environment, the detection time is long, and the extraction reagents used contain toxic and harmful substances, which pose a safety hazard. By adjusting the pH value of the lysate, the extraction of DNA and RNA can be achieved separately, which solves the problem of existing kits, etc. The problem that the method can only extract a single nucleic acid
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 2
[0030] (1) Prepare the lysate, accurately weigh 29.54g of guanidine isothiocyanate, 1.67ml of sodium citrate of 0.75mol / L and 2.5ml of 10% sodium lauryl sulfate, add 10ml of deionized water, heat and stir to dissolve, use hydrochloric acid to adjust The pH value of solution is 7, is mixed with final concentration and is the guanidine isothiocyanate of 5mol / L, the sodium citrate of 25mol / L and the sodium lauryl sulfate solution of 0.2%, be settled to 50ml, put into the brown bottle, Store at room temperature;
[0031] (2) Get 4 parts of foot-and-mouth disease virus and Peste des Petits Ruminants virus culture fluid respectively 200uL, add the lysate of 1ml preparation respectively;
[0032] (3) Place at room temperature for 10 minutes, centrifuge at 10,000 g for 5 minutes, and transfer the supernatant to a new tube;
[0033] (4) Add 0.5ml of isopropanol to the supernatant, centrifuge at 10000g for 8 minutes, discard the supernatant, and the precipitate is RNA;
[0034] (5) Us...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
Login to View More Abstract
The invention discloses a quick extraction method for nucleic acid of diseased material of animal tissues, and relates to the technical field of nucleic acid extraction. The quick extraction method comprises the following steps of (1) preparation of lysate; (2) cracking of tissues; (3) primary separation; (4) precipitation of nucleic acid; (5) purification of nucleic acid. The quick extraction method has the advantages that by improving the used lysate in the nucleic acid extraction process, the nucleic acid extraction method is greatly simplified, the operation is simple, convenient and safe,the cost is low, and the problems of failure to adapt to outdoor environments and longer detection time in the existing nucleic acid extraction method and safety hidden hazards due to containing of poisonous and harmful matters in the used extraction reagent are solved; by adjusting the pH (potential of hydrogen) value of the lysate, the DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) canbe respectively extracted, so as to solve the problem of only extraction of single nucleic acid in the existing kit method and the like.
Description
technical field [0001] The invention belongs to the technical field of nucleic acid extraction, in particular to a rapid extraction method for nucleic acid from animal tissue disease materials. Background technique [0002] Nucleic acid (including DNA and RNA) extraction and purification is a basic operation in molecular biology. Molecular biology technology has been more and more used in the detection and diagnosis of animal diseases due to its advantages of rapidity, sensitivity and specificity. At present, nucleic acid extraction includes phenol extraction method, salt washing method, TRIZOL method, magnetic bead method and kit method, etc., but the traditional detection methods have high requirements on instruments and environment, and cannot meet outdoor operations. In order to further improve port animal The level of quarantine, it is particularly important to invent a new method that can meet both the rapid detection technology of the port and the precise detection te...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
Login to View More Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 李林红
Owner 芜湖帮许来诺医疗设备科技有限公司