KR-20260066548-A - Method for manufacturing nucleic acid selection solution and nucleic acid selection solution prepared thereby

Abstract

A method for preparing a nucleic acid screening solution and a nucleic acid screening solution prepared thereby are disclosed. Magnetic particles are dispersed in distilled water using ultrasound, and a 1.0x adsorption solution is prepared by mixing the distilled water in which the magnetic particles are dispersed with a 1.2x adsorption solution.

Inventors

• 김현
• 하정미
• 주재현

Assignees

• 주식회사 나노실리칸첨단소재

Dates

Publication Date

20260512

Application Date

20241104

Claims (6)

1. A step of dispersing magnetic particles in distilled water using ultrasound; and A method for preparing a nucleic acid screening solution characterized by including the step of preparing a 1.0x adsorption solution by mixing distilled water in which the magnetic particles are dispersed with a 1.2x adsorption solution.
2. In Article 1, A method for preparing a nucleic acid screening solution characterized in that the above 1.2x adsorption solution comprises polyethylene glycol and a salt.
3. In Paragraph 2, A method for preparing a nucleic acid screening solution characterized in that the above salt is NaCl.
4. In Paragraph 2, A method for preparing a nucleic acid screening solution characterized in that the viscosity of the above 1.0x adsorption solution is 1 or higher and 50 or lower.
5. In Article 1, A method for preparing a nucleic acid screening solution characterized in that the degree of dispersion of the magnetic particles in the above 1.0x adsorption solution is PDI 0.25 or less.
6. A nucleic acid screening solution prepared by the method for preparing an adsorption solution described in claim 1.

Description

Method for manufacturing nucleic acid selection solution and nucleic acid selection solution prepared thereby An embodiment of the present invention relates to a method for preparing a nucleic acid selection solution for selecting nucleic acids of a desired size and a nucleic acid selection solution prepared thereby. This invention was carried out as part of Pyeongtaek City's '2024 Pyeongtaek City Hidden Champion Development Support Project' (Research Management Agency: Gyeonggi Economic & Science Promotion Agency (GBSA), Research Project Title: Development and Mass Production for Localization of Size Selection Beads for Next-Generation Sequencing (NGS), Lead Agency: Gyeonggi Economic & Science Promotion Agency (GBSA)). Nucleic acid capture or screening methods using magnetic beads are being utilized in various nucleic acid-based research. In particular, nucleic acid size screening is a preliminary step for Next-Generation Sequencing (NGS), aiming to selectively extract nucleic acids of an appropriate size based on the target genetic material. Since nucleic acids of various sizes are mixed within a sample, selecting those of the desired size can significantly improve the efficiency of subsequent sequencing. However, the adsorption solutions used to bind nucleic acids of a desired size to the surface of magnetic particles typically have high viscosity and concentration, which leads to the problem of low dispersibility of the magnetic particles. This low dispersibility of magnetic particles results in a reduction in the surface area available for nucleic acid binding, thereby lowering the nucleic acid extraction efficiency. In particular, since small nucleic acids have relatively lower adsorption capacity than large nucleic acids, the dispersibility of magnetic particles significantly affects the extraction yield when attempting to select small nucleic acids. FIG. 1 is a diagram illustrating an example of the dispersibility of magnetic particles in an adsorption solution. FIG. 2 is a flowchart illustrating an example of a method for preparing a nucleic acid screening solution using a magnet, FIG. 3 is a diagram illustrating an example of a method for preparing a nucleic acid screening solution according to an embodiment of the present invention, and, Figure 4 is a diagram showing the results of nucleic acid screening experiments using a nucleic acid screening solution prepared by the nucleic acid screening solution preparation method of Figures 2 and 3. Hereinafter, a nucleic acid screening solution and a method for manufacturing the same according to an embodiment of the present invention will be examined in detail with reference to the attached drawings. Figure 1 is a diagram illustrating an example of the dispersibility of magnetic particles in an adsorption solution. Referring to FIG. 1, the nucleic acid screening solution (100, 150) includes an adsorption solution (110, 160) and magnetic particles (120, 170). This embodiment illustrates a case where nucleic acids (130, 180) are dissolved together in the nucleic acid screening solution (100, 150) to aid in understanding nucleic acid detection using magnetic particles (120, 170). Since the magnetic particles (120, 170) for nucleic acid screening themselves are of a widely known composition, further description of the magnetic particles (120, 170) is omitted. Various existing types of magnetic particles (120, 170) for nucleic acid extraction may be used in this embodiment. The adsorption solution (110, 160) contains polyethylene glycol (PEG) and a salt (e.g., NaCl). The polyethylene glycol and the salt serve to bind nucleic acids (130, 180) dissolved in the adsorption solution (110, 160) to the surface of magnetic particles (120, 170). Depending on the concentration of polyethylene glycol and/or salt in the adsorption solution (110, 160), the size of the nucleic acids (130, 180) adsorbed onto the magnetic particles (120, 170) can be adjusted. The extraction efficiency of nucleic acids (130, 180) is affected by the dispersibility of magnetic particles (120, 170) in the adsorption solution (110, 160). When the dispersibility of magnetic particles (120, 170) in the adsorption solution (110, 160) is low (100), the total surface area of the magnetic particles to which nucleic acids (130, 180) can bind is smaller compared to when the dispersibility is high (150), and as a result, the nucleic acid detection efficiency is lowered. Since polyethylene glycol in the adsorption solution (110, 160) has viscosity, there is a problem in that if the concentration of polyethylene glycol is increased to detect nucleic acids of a desired size, the dispersibility of the magnetic particles becomes lower, and as a result, the extraction efficiency also decreases. Figure 2 is a flowchart illustrating an example of a method for preparing a nucleic acid screening solution using a magnet. Referring to FIG. 2, magnetic particles are mixed with distilled water, and then ultrasound is