The present disclosure discloses a method for preparing a spatial
chip, a spatial
chip, and a use thereof. The method comprises: hybridizing to attach a single-stranded
nucleic acid molecule to a surface of a
solid-phase substrate, the single-stranded
nucleic acid molecule comprising a first
fixed sequence, a spatial
barcode sequence, a sequencing primer binding sequence and a second
fixed sequence, a first probe being attached to the surface of the
solid-phase substrate of the
chip, and the first probe comprising a complementary sequence of the first
fixed sequence and a complementary sequence of the second fixed sequence; ring closing, causing the single-stranded
nucleic acid molecule to form a circular
nucleic acid molecule; rolling circle amplification; sequencing, to obtain sequence information of the spatial
barcode sequence; synthesizing a single-stranded
nucleic acid molecule by using the rolling circle amplification product as a template, to obtain a single-stranded
nucleic acid molecule library; and fixing and clustering, fixing the single-stranded nucleic
acid molecule library on the surface of the
solid-phase substrate to form a single-stranded nucleic
acid molecule cluster. According to the present disclosure, a chip having a nucleic acid probe carrying spatial position coordinate information can be prepared.