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Amphibian skins have been used for centuries as components of traditional medicines due to their possession of a wide spectrum of pharmacological effects.In recent years,analytical technologies have shown that they contain a plethora of active compounds,especially peptides.As amphibians are currently in global decline,it was the aim of our research to critically evaluate several methods oftissue/skin secretion preparation that would permit subsequent parallelpeptidome and transcriptome analyses,preferably without recourse to sacrifice of the donor specimen.cDNA libraries were constructed from both lyophilised venom and macerated air-dried skin using magnetic oligo—dTbead-captured P olyadenylated mRNA as templates.Those CDNAs encoding skin gra nular gland—derived pe ptideprecursors were amplified by a RACE PCR strategy.Products from different species of amphibian were purified,clonedand sequenced.Subsequent to prediction ofm ature P eptide amino a cid sequences from re spective cloned pr ecursorcDNAs,the presence of each peptide in the original preparation(skin secretion or dried skin)from each amphibian specieswas confirmed by a combination ofreverse—phase HPLC fractionation,MALDI-TOF mass spectrometry and automatedEdman degradation microsequencing.