To obtain full-length genes related to dormancy release and provide resources for tree peony genomic, flower buds of different chilling duration was used to isolate mRNA in tree peony. A normalized full-length yeast hybridization cDNA library was then constructed by DSN (duplex- specific nuclease) normalization method combined with SMART (switching mechanism at 5' end of RNA transcript) technique. The amplified cDNA was recombined with pGADT7-Rec, and transformed to yeast AH109. The titer of un-amplified cDNA library was about 1.77×106 cfu/ml, and contained 2.3×108 independent clones. The abundance of transcripts 18S rRNA decreased 26 in normalized cDNA library comparing with that in non-normalized samples detected by PCR and Tanon GIS analysis. The average size of cDNA inserts was 1200 bp with a recombination rate of over 95%. These results indicated that the normalized yeast hybridization cDNA library has been successfully established with high quality, which is convenient to screen transcription factors and interaction protein regulated by chilling temperature, and finally to develop gene regulation network during dormancy release in tree peony.