Molecular Ecology Notes (2005) 5, 654–656 doi: 10.1111/j.1471-8286.2005.01023.x © 2005 Blackwell Publishing Ltd Blackwell Publishing, Ltd. PRIMER NOTE Characterization of microsatellite loci in the brown treecreeper (Climacteris picumnus) and cross-species amplification in the white-throated treecreeper (Cormobates leucophaeus) ERIK D. DOERR*† * Program in EEC Biology, University of Nevada-Reno, Reno, NV 89557 USA, † School of Botany and Zoology, Australian National University Canberra, ACT 0200 Australia Abstract Eight polymorphic microsatellite markers were developed for the brown treecreeper, Cli- macteris picumnus. The number of alleles ranged from three to 25 per locus with observed heterozygosities between 0.05 and 0.76. Seven of the eight primer pairs also amplified poly- morphic microsatellite loci in the white-throated treecreeper (Cormobates leucophaeus). These markers are likely to be useful for population genetic and parentage studies in any of the Australasian treecreepers (Climacteridae) and are the first genetic markers developed for any member of this passerine family. Keywords: Climacteridae, Climacteris picumnus, Cormobates leucophaeus, microsatellites, primers, treecreepers Received 26 January 2005; revision received 3 March 2005; accepted 1 April 2005 The Australasian treecreepers (Climacteridae) are one of the oldest passerine families. The brown treecreeper (Climacteris picumnus) is typical of the family in being highly social, exhibiting not only cooperative breeding but also complex associations between breeding groups (Doerr & Doerr 2001b). The white-throated treecreeper ( Cormobates leucophaeus ) is ecologically similar, but breeds only in pairs (Doerr & Doerr 2001a). I developed microsatellite markers (the first for any climacterid species) in order to investigate patterns of paternity and the population genetics of these two species as part of a comparative study. Genomic DNA was isolated from the blood of a female brown treecreeper via phenol–chloroform extraction (Sambrook et al . 1989). Approximately 6 μ g of this DNA was partially digested with Hae III and Alu I. Fragments between 220 and 1000 bp were isolated by electrophoresis and purification using a BRESAclean kit (Geneworks). Frag- ments were ligated into Sma I-digested and dephosphory- lated pUC18 (Qbiogene), and the plasmids transformed into Escherichia coli cells (TOP10 One Shot chemically competent cells, Invitrogen). Cells were spread on LB-agar containing 100 μ g/mL ampicillin and incubated overnight, producing a library of about 21 500 recombinant colonies. An aliquot of this library was spread on LB-agar contain- ing ampicillin and resulting colonies blotted onto nylon membranes (Hybond-N, Amersham). After UV cross-linking, membranes were probed with oligonucleotides [(AC) 15 , (AG) 15 , (CAAA) 8 , (AAAG) 8 ] that were end-labelled with [ γ 32 P] dATP. Eighty-nine positive clones were picked, replated and probed a second time, yielding 28 double pos- itives. A separate aliquot of the library was plated out and probed with the same oligonucleotides as previously plus all possible trimeric repeats [(AAC) 10 , (AAG) 10 , etc.], yield- ing an additional 47 positive and 29 double-positive clones. Insert DNA from each of the 57 double positives was amplified by polymerase chain reaction (PCR) with universal M13 primers and gel purified as above. Purified clones were sequenced in both directions using M13 primers and a BigDye Terminator cycle sequencing kit (Applied Biosys- tems). Products were separated on an ABI PRISM 377 DNA sequencer (Applied Biosystems), and sequences aligned using sequencher 3.0 (Gene Codes Corporation). Fifteen clones had microsatellite regions at least 20 bp Correspondence: E. D. Doerr, School of Environmental Sciences and Natural Resources Management — Zoology, University of New England, Armidale, NSW 2351, Australia. Fax: +61-2-6773-3814; E-mail: erik.doer@anu.edu.au