The invention discloses a multi-
gene repetitive sequence sequencing method based on
fluorescence imaging, and belongs to the technical field of
gene sequencing. The method comprises the following steps: sample targeting pretreatment, fluorescent probe synthesis, in-situ
hybridization reaction, multi-layer
fluorescence imaging acquisition,
fluorescence signal analysis and sequence recognition, and sequence
verification and splicing, and the sample targeting pretreatment is subjected to
microdissection,
enzyme digestion and
magnetic bead enrichment to obtain high-purity and high-concentration target
DNA. The design of the fluorescent probe is guaranteed by multiple groups of fluorescent labels and high purity, in-situ hybridization enhances the combination efficiency by optimizing fixing and washing conditions, a light path and an imaging interval are accurately controlled by multi-layer
fluorescent imaging, a high-resolution and low-interference image is obtained, complete
signal information is reserved, accurate conversion from the image to a sequence is realized by fluorescent
signal analysis, and the detection accuracy is improved. Sequence
verification splicing is subjected to secondary hybridization
verification and overlapping region integration,
short segment limitation is broken through, the final sequence accuracy is improved, and the multi-
gene repetitive sequence sequencing requirement is comprehensively met.