Capturing method of specific target RecA (Right External Carotid Artery) mediated DNA (Deoxyribose Nucleic Acid) fragments

Abstract

The invention belongs to the technical field of biology and biochemistry, and relates to a method for specifically capturing RecA (Right External Carotid Artery) mediated homologous target DNA (Deoxyribose Nucleic Acid) fragments in samples. The method comprises the following steps of: performing an in-vitro simulation on an action mode of the RecA in in-vivo homologous recombination; designing homologous probes aiming at the target DNA fragments; and specifically capturing the homologous target DNA fragments in the samples in a reaction system containing the RecA and ATP-Gamma-S through RecA assisting probes and an elution reagent. The method can be used for the capture sequencing of exons or other target fragments. Thus, a technology for detecting the rare mutation sites of tumors and other diseases is improved. By utilizing the method, the problems in the prior art that the uniformity of Tm values of the probes designed for capturing the exons or the other target fragments is required, the specificity is not generated in probe hybridization, the initial quantity of the required samples is relatively large and the like are solved. The method also can be used for capturing in-vivo DNA binding proteins based on the homologous recombination of Rad51 under the state of eukaryotic chromatins.

Description

technical field [0001] The invention belongs to the technical field of molecular biology-biochemistry, and relates to a method for capturing homologous target DNA fragments in samples, in particular to a method for specifically capturing homologous target DNA fragments in samples under the mediation of RecA. This method can be used for capture sequencing of exons or other purpose fragments, and provides technical assistance for the detection of rare mutation sites in tumors and other diseases. Background technique [0002] Studies have shown that exome sequencing provides a more effective technical means for disease detection and mechanism research. Compared with traditional human whole genome sequencing, exome sequencing can detect new disease-causing genes, especially rare gene variants. , showing a high detection efficiency. It has been reported that exome sequencing can identify a series of rare single nucleotide polymorphisms (SNPs), DNA missense mutations, and inserti...

AI Technical Summary

Problems solved by technology

However, the high cost and high time consumption of exome sequencing are still important factors that limit its large-scale application, and the probe hybridization transaction is non-specific during exome sequencing, and the Tm value should be considered in the design of the probe to make it uniform. The coverage degree, the initial amount of sample required is relatively large, and many factors seriously limit its large-scale application in clinical disease research.

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