A method for DNA fragmentation and a device for realizing the method

Abstract

The invention discloses a DNA fragmentation method and a device for realizing the method. Bubbles are continuously bubbled in a DNA solution, and the instantaneous fluid shear force generated during the bubble bursting acts on the DNA molecules in the solution. When the shear force is greater than the intermolecular force in the DNA molecular chain, the DNA molecule will be sheared and broken, thereby achieving the fragmentation of the DNA molecule. The volume of the DNA sample required by the present invention is small, the experimental process is easy to control, the required device is simple, the cost is low, the operation is simple, it is not easy to be blocked, no high pressure is required, a large amount of dead volume will not occur, and the sample will not be polluted. The random shearing of DNA has the advantages of relatively controllable fragment length and small dispersion of fragment length, and the DNA sheared fragments are evenly distributed within the predetermined size.

Description

technical field [0001] The invention relates to a method for fragmenting DNA and a device for realizing the method, in particular to a method for fragmenting DNA by using air bubbles to cut DNA, with controllable length of DNA fragments and random fragmentation of DNA fragments. Background technique [0002] Randomly distributed DNA fragments are one of the key steps for technologies such as gene sequencing and DNA microarray-based analysis. For genetic analysis, the range of DNA fragment lengths is usually chosen to approximate the average length of a single gene in related organisms, or based on methods that automatically generate sequence data for entire databases. In molecular diagnosis, different detection methods are used to screen some genes from a sample, and the target gene is hybridized with a complementary probe molecule. Therefore, for molecular diagnosis applications, DNA fragmentation is an important sample pretreatment step, short DNA fragments are essential ...

AI Technical Summary

Problems solved by technology

The advantage of the restriction endonuclease method is that DNA can be cut at a specific position to obtain fragments; however, it is prone to sample contamination, sample degradation, large dispersion of the length of the generated DNA fragments, and a relatively large number of single-stranded fragments. fracture

The sonication method uses sound waves to generate a shear force, which can lead to the breakage of long DNA strands. This method is fast and simple; but it will cause damage to the DNA sample, and the resulting fragment lengths have a large dispersion and are difficult to integrate.

The jet atomization method uses a pressurized nozzle to generate a mechanical shear force to shear the DNA sample, which is fast and efficient; however, the equipment is expensive and requires a large amount of DNA sample, which is difficult to integrate into the device

Fragmented DNA can be generated based on the hydrodynamic shearing DNA fragmentation method, and the length dispersion of the obtained DNA fragments is small. This method can obtain DNA fragments with double-strand breaks, which are random breaks and are easy to integrate into devices; The disadvantage is that the experimental system requires high pressure, the device preparation is relatively expensive, there is a large amount of dead volume, and the microstructure device is easy to block, which is not suitable for miniaturization and integration

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