Background: U6 snRNA, as a component of the spliceosomes, is involved in splicing of pre-mRNAs.There is no report about U6 in chicken.Methods: in this study, a 94-bp fragment of chicken U6 snRNA was amplified by reverse transcription PCR method, then cloned and sequenced.In order to get full-length U6 snRNA in chicken, by using the 94-bp sequence as a probe, we screened the chicken genomic sequence database.Results: we determined the chicken full-length U6 snRNA sequence was probably 106-bp long.The sequence bioinformatics analysis showed chicken U6 snRNA had 100% sequence identity to its human counterpart.We identified four copies of chicken U6 snRNA gene in chicken genome.Three of them are clustered in a 2-kb DNA region in chromosome 28, the other is in chromosome 18.The 5-flanking regions of the four copies of chicken U6 snRNA have been validated to have promoter activity in vitro by other people.So we conclude that the four identified U6 RNA genes are real genes.Promoter sequence analysis showed that the four identified chicken U6 promoters contained common RNA Pol Ⅲ promoter elements: distal sequence element (DSE), proximal sequence element (PSE) and TATA-box like element.Conclusions: we obtained a full-length chicken U6 snRNA, and determined its copy number successfully.Our data lay a firm foundation for chicken miRNA quantification and U6 snRNA transcriptional regulation .