Nucleic acid fragment sorting and purifying reagent and method

Abstract

The invention discloses a nucleic acid fragment sorting and purifying reagent and method. The reagent comprises a magnetic bead binding solution, a washing solution and an eluting solution, and can beused for sorting and purifying targeted nucleic acid fragments of different segments in a mixture sample. Sorting of the targeted nucleic acid fragments comprises the following steps that a first nucleic acid mixture containing different segments and the magnetic bead binding solution are mixed uniformly, placed at the room temperature, and then placed on a magnetic stand for standing, supernatant is discarded, and washing and elution are carried out; the volume mixing ratio of the first nucleic acid mixture to the magnetic bead binding solution is 1:(1.2-0.45). In addition, whether or not secondary sorting is performed can be selected according to requirements, that is to say, the first nucleic acid mixture and magnetic beads are combined with the separated supernatant to be taken as a second nucleic acid mixture, and then binding separation, washing and elution are carried out again; the targeted nucleic acid fragments of different segments can be sorted by adjusting the volume of the added magnetic bead binding solution. The provided reagent and method can be used for downstream nucleic acid fragment analysis, second generation sequencing platform library construction and the like.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to a reagent and method for sorting and purifying nucleic acid fragments. Background technique [0002] The British biochemist Sanger invented the hydrogen-oxygen-terminated DNA sequencing method, which greatly promoted the industry prospect of gene sequencing development, and then the second-generation sequencing technology and the third-generation sequencing technology gradually appeared, and these two technologies are generally referred to as next-generation sequencing. Technology (NGS), next-generation sequencing has a wide range of applications, as follows: 1. Denovo sequencing, resequencing and other DNA sequencing; 2. RNA sequencing; 3. Metagenome sequencing; 4. Epigenetics. The nucleic acid templates required by next-generation sequencing have certain requirements in terms of quality and quantity, and the fragment size of nucleic acid templates is of great significance for...

AI Technical Summary

Problems solved by technology

[0004] At present, there are two methods for sorting target nucleic acid fragments of different sizes, one is recovery by gel cutting, but the experimental method is cumbersome and time-consuming, and there is a certain risk in ultraviolet irradiation, and it is easy to cause mutations in nucleic acid bases

Another method is described in the patent US6534262 to use polyethylene glycol (PEG) and sodium chloride to separate and precipitate nucleic acids, and effectively sort out target nucleic acid fragments, but it takes a long time to process, the operation is complicated, and the nucleic acid content in the nucleic acid mixture When higher, there will be good recovery efficiency

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