The invention relates to the technical field of high throughput sequencing, and discloses a method for establishing a DNA (Deoxyribose Nucleic Acid) library based on an illumina sequencing platform. The method comprises the following steps: (1) randomly interrupting the genome DNA; (2) filling the tail ends, and adding biotin labeled cyclizing connectors at two ends; (3) separating DNA fragments, wherein the sequence of the step (2) and the step (3) can be changed;; (4) cyclizing, and removing uncyclized DNA fragments; (5) randomly interrupting cyclized DNA; (6) separating biotin labeled DNA fragments; (7) filling the tail ends of the biotin labeled DNA fragments; (8) adding an A alkali group; (9) connecting with the sequencing connectors; (10) performing PCR amplification and enrichment, thereby obtaining the sequencing library. The method adopts a Cre recombinase system to cyclize the DNA fragments, the cyclizing only takes 1 hour, the time for establishing the library is greatly shortened, the large-fragment DNA library is successfully established, and splicing of genomes in sequencing can meet the standard of a fine diagram or accomplished diagram.