The invention provides a sequencing method based on a
gene capture technology. The sequencing method has the principle that a
recombinase compound is used for assisting a single-stranded
DNA probe with a closed 3' end to efficiently and specifically hybridize into a double-stranded target
DNA molecule under the condition of 37 DEG C, and then an
affinity label on the single-stranded
DNA probe is used for carrying out specific enrichment on the target DNA. The method not only has the advantages of relatively strong long probe
pairing, low probe design requirement and the like of a probe
hybridization capture technology, but also has the advantages of simplicity in operation, short time consumption, no need of
thermal denaturation of DNA, high capture efficiency, low cost and the like of a
CRISPR / dCas9
system, and the probe does not participate in subsequent amplification, so that the identification accuracy is improved. Therefore, compared with other capture technologies, the RATE-seq has the obvious advantages of simplicity in operation, high capture efficiency, extremely short consumed time (30 minutes), good stability, low cost and the like, and is very suitable for
gene capture and automatic application. In addition, the RATE-seq can effectively separate and remove human host DNA in a
pathological sample, and the
detection rate of pathogenic microorganisms is increased.