Trace nucleic acid sequencing pretreatment method based on magnetic bead coating and kit

Abstract

The invention discloses a trace nucleic acid sequencing pretreatment method based on magnetic bead coating and a kit. The trace nucleic acid sequencing pretreatment method based on magnetic bead coating comprises the following steps of: firstly coating DNA (deoxyribonucleic acid) to be treated with magnetic beads, carrying out tail end repairing on the DNA to be treated, which is coated with the magnetic beads, purifying, leaving the magnetic beads, carrying out A addition reaction, purifying again, leaving the magnetic beads, carrying out joining reaction, purifying, discarding the magnetic beads, carrying out PCR (polymerase chain reaction), and purifying by adding the magnetic beads after completing polymerase chain reaction to obtain the treated DNA. The trace nucleic acid sequencing pretreatment method based on magnetic bead coating has the beneficial effects that the frequency of transferring trace nucleic acids in a sequencing pretreatment process is effectively reduced by coating the DNA with the magnetic beads, thereby greatly reducing the loss of the nucleic acids in a sequencing pretreatment process; reaction conditions are mild; meanwhile, the DNA damage is also reduced; the success rate of trace nucleic acid sequencing pretreatment is increased to 90% or above; the method has the characteristics of high success rate, high quality and low initial amount in comparison with a traditional nucleic acid sequencing pretreatment method; and used reagent raw materials are low in cost, simple and easily available.

Description

technical field [0001] The present invention relates to the technical field of molecular biology, more specifically, to the technical field of DNA, in particular to a pretreatment method and kit for micro nucleic acid sequencing based on magnetic bead coating. Background technique [0002] DNA is the material basis of human inheritance. In order to clarify the function and structure of DNA in the process of human inheritance and development, the detection of DNA sequence has become the most important means of studying DNA. In 1977, Sanger invented the landmark terminal termination sequencing method, and in the same year A.M.Maxam and W.Gilbert invented the chemical degradation method. The Sanger method has become the mainstream of the first-generation DNA sequencing because it is simple and fast, and has been continuously improved. With the continuous innovation of science and technology, the first-generation DNA sequencing technology (Sanger) can no longer meet the needs o...

AI Technical Summary

Problems solved by technology

[0003] The next-generation sequencer itself cannot directly read the DNA sequence, and the DNA needs to be homogenized before being placed in the sequencer to be recognized by it.

[0004] As the next-generation sequencing technology continues to penetrate the medical field, more and more clinical samples are used for next-generation sequencing. However, clinical samples are very precious and the amount of DNA is scarce. It is difficult to achieve the total amount of DNA required for classic nucleic acid sequencing pre-processing. Therefore, it has become a bottleneck problem for the application of this technology to clinical samples.

Images