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CN-121975793-A - Method for extracting DNA from cotton in high flux

CN121975793ACN 121975793 ACN121975793 ACN 121975793ACN-121975793-A

Abstract

The invention discloses a method for extracting DNA from cotton in high flux, belonging to the field of plant biotechnology. The invention provides a lysis buffer solution for high-throughput DNA extraction of cotton, which comprises the following components of 50-80 g/L, naCl 2-4 g/L, SDS of NaOH, 0.1 g/L, EDTA-2 Na.2H2H 2 O, 0.8-1.5 g/L, PVP, 10-30 g/L of gelatin 2g/L and 8-12 g/L of ascorbic acid, wherein the components are dissolved in deionized water. The high-throughput DNA extraction method for cotton tissues can realize high-efficiency cotton tissue DNA extraction, greatly simplify the extraction steps, realize high-throughput extraction in a laboratory by a single person, efficiently obtain DNA for PCR amplification, provide reliable sample support for subsequent molecular biological research, and provide a convenient tool for scientific researchers.

Inventors

  • LIU SHIYU
  • CUI YANLI
  • WANG JUAN
  • CHEN WEI
  • FENG TIANYU
  • LI JING

Assignees

  • 中国农业科学院棉花研究所

Dates

Publication Date
20260505
Application Date
20260206

Claims (5)

  1. 1. A lysis buffer solution for high-throughput DNA extraction of cotton is characterized by comprising the following components of 50-80 g/L, naCl 2-4 g/L, SDS of NaOH, 0.1 g/L, EDTA-2 Na.2H2H 2 O0.8-1.5 g/L, PVP, 10-30 g/L of gelatin 2 g/L and 8-12 g/L of ascorbic acid, wherein the components are dissolved in deionized water.
  2. 2. The lysis buffer of claim 1, comprising the components of NaOH 50g/L, naCl 3g/L, SDS 0.1 g/L, EDTA-2 Na.2H2H 2 O1 g/L, PVP 20 g/L, gelatin 2 g/L and ascorbic acid 10 g/L.
  3. 3. A method for extracting DNA from cotton in high flux, which is characterized by comprising the following steps: (1) Mixing cotton tissue samples, grinding beads and the lysis buffer of claim 1 or 2 for mechanical disruption; (2) Standing and incubating the crushed mixture at room temperature to lyse cells and release DNA; (3) Sucking the supernatant obtained in the step (2), and diluting with sterile water to obtain cotton DNA.
  4. 4. The method of claim 3, wherein the incubation time is 10-20 minutes.
  5. 5. A kit for high throughput DNA extraction of cotton comprising the lysis buffer of claim 1 or 2.

Description

Method for extracting DNA from cotton in high flux Technical Field The invention relates to the technical field of plant biology, in particular to a method for extracting DNA (deoxyribonucleic acid) from cotton at high flux. Background In plant molecular biology research, DNA extraction is a fundamental step in performing genomic analysis, genetic improvement, and crop breeding. The current common cotton DNA extraction method mainly comprises a traditional CTAB method and a commercial kit extraction method. These methods, while effective, still suffer from the following disadvantages: 1. The traditional CTAB method has a large number of steps, and relates to the processes of tissue disruption, warm bath, separation, purification, cleaning, drying, re-dissolution and the like, and the method generally takes more than 2 hours, is complex in operation, has large dependence on instruments and is difficult to adapt to the high-flux extraction requirement. 2. Complicated operation, high labor intensity, complex flow, higher requirement on the experimental skill of operators and increased burden of the experimenters. 3. The environmental friendliness is insufficient, and the organic solvents such as phenol, chloroform and the like used in the CTAB method not only pollute the environment, but also form potential risks for the health of operators. 4. The cost is high, and the commercial kit (such as column extraction of about 4 yuan/sample and magnetic bead method of about 5 yuan/sample) on the market at present is not economical for conventional experiments such as positive seedling identification, mutation detection and the like which are carried out in a large amount in a laboratory, although the quality of the extracted DNA is high, and the method is suitable for experiments with strict requirements on purity such as re-sequencing. Disclosure of Invention The invention aims to provide a method for extracting DNA from cotton in high flux so as to solve the problems in the prior art. The invention relates to a DNA extraction technology in cotton genetics and molecular biology, which is mainly applied to the fields of cotton genome research, breeding and the like. Aiming at the problems in the prior art, the invention establishes a high-flux DNA extraction method suitable for cotton tissues by simplifying steps, shortening time and improving DNA extraction yield, and provides more efficient and reliable technical support for plant genetic research. The invention aims to solve the following specific technical problems: 1. solves the problems of complicated steps, long time consumption and the like of the traditional CTAB method, and establishes a high-throughput extraction flow which is simpler in operation and can finish mass sample processing in a shorter time. 2. Solves the problems of high labor intensity and high dependence on operation skills of the existing method, and provides an extraction scheme which is easier to standardize and can reduce the complexity of manual operation. 3. Improving the safety and environmental risks of organic solvents used in traditional methods, a more environmentally friendly and operator friendly alternative is sought. 4. On the premise of meeting the requirements of conventional molecular detection (such as positive seedling identification and mutation detection), the cost of the single-time extracted reagent and consumable materials is obviously reduced so as to meet the requirements of large-scale sample detection in a laboratory. The invention has the core task of providing a high-flux DNA extraction method which is suitable for cotton organization, has the characteristics of rapidness, low cost, simple and convenient operation and environmental friendliness. In order to achieve the above object, the present invention provides the following solutions: According to one of the technical schemes, the lysis buffer solution for high-throughput DNA extraction of cotton comprises the following components of 50-80 g/L, naCl 2-4 g/L, SDS 0.1 g/L, EDTA-2 Na.2H2O 0.8-1.5 g/L, PVP 10-30 g/L, 2 g/L of gelatin and 8-12 g/L of ascorbic acid, wherein the components are dissolved in deionized water. The second technical scheme of the invention is a method for extracting DNA from cotton in high flux, which comprises the following steps: (1) Mixing cotton tissue samples, grinding beads and the lysis buffer solution, and mechanically crushing; (2) Standing and incubating the crushed mixture at room temperature to lyse cells and release DNA; (3) Sucking the supernatant obtained in the step (2), and diluting with sterile water to obtain cotton DNA. In a third technical scheme of the invention, a kit for high-throughput DNA extraction of cotton comprises the lysis buffer. Based on the technical scheme, the invention has the following technical effects: The high-throughput DNA extraction method for cotton tissues can realize high-efficiency cotton tissue DNA extraction, greatly simplify the extract