The objective of our study is to obtain a high-quality reference genome of Poeciliopsis prolifica with gene annotation and identify the exonic SNP in inbred lines by RNA sequencing to provide the essential genetic toolkit for evolutionary and comparative genomic studies for P. prolifica and facilitate the reseach of placental evolution. Overall design: Whole-body of male P. prolifica was used for high molecular weight genomic DNA extraction and genome sequencing and the juvenile whole-body fish from line A (n=3) and line B (n=3) were used for RNA sequencing.