The　defensive　skin　secretions　of　amphibians　are　a　rich　source　of　bioactive　peptides.　Here　we　describe　a　rapid　technique　for　skin　granular　gland　transcriptome　cloning　from　a　surrogate　tissue-the　secretion　itself.　cDNA　libraries　were　constructed　from　lyophilized　skin　secretion　from　each　of　the　Chinese　frogs　(Rana　schmackeri,　Rana　versabilis,　and　Rana　plancyi　fukienensis)　using　magnetic　oligo(dT)　bead-captured　polyadenylated　mRNA　as　templates.　Specific　esculentin　cDNAs　were　amplified　by　3′-RACE　using　a　degenerate　primer　designed　for　a　consensus　nucleotide　sequence　in　the　5′　untranslated　region　of　previously　characterized　ranid　frog　peptide　cDNAs.　The　cloned　cDNAs　were　found　to　encode　the　antimicrobial　peptides　esculentins　1　and　2　from　each　of　the　species　examined.　The　presence　of　predicted　peptide　structures　in　skin　secretions　was　confirmed　by　MALDI-TOF　mass　spectrometry　and　automated　Edman　degradation.　This　experimental　approach　can　thus　rapidly　expedite　parallel　transcriptome　and　peptidome　analysis　of　amphibian　granular　gland　secretions　without　harming　or　sacrificing　donor　animals.　©　2005　Elsevier　Inc.　All　rights　reserved.