The preparation of sex-chromosome-specific painting probes and construction of sex chromosome DNA library in half-smooth tongue sole (Cynoglossus semilaevis)

An improved method of slide preparation on special membrane was developed and a high quality of chromosome spreads was obtained from female half-smooth tongue sole (Cynoglossus semilaevis). The W chromosome was isolated and the sex-chromosome-specific painting probes were prepared by means of laser microdissection. The hybridization signals revealed by Cot-1 blocked and unblocked slides indicated the existence of repetitive DNA in W and putative Z chromosomes. A W chromosome library containing 596 clones was constructed and sequences were analyzed using BLAST and repetitive sequences analysis. The sex-chromosome-specific painting probes and the W chromosome library we prepared in this study will be great help for better understanding of the C. semilaevis W chromosome.