Ab 3730 dna analyser

DNA sequence variation in the hypervariable Domain I of the mtDNA CR was determined using marsupial-specific primers [46 (link)] and single strand conformation polymorphism (SSCP [47 (link)]) as previously described [48 ]. All unique haplotypes were sequenced including multiple representations (up to four where available) using BigDye termination (Perkin-Elmer Applied Biosystems, Norwalk, CT, USA) and resolved in an AB 3730 DNA Analyser (Applied Biosystems, USA) at the Ramaciotti Centre, UNSW or on an ABI 377 Sequencer at Macquarie University. Sequences were edited using Sequencher v4.8 (Gene Codes Corporation, Ann Arbor, USA) and aligned using Clustal X [49 ].

Individuals were genotyped at 16 polymorphic autosomal microsatellite loci derived from the tammar wallaby (T3.1T, T15.1, T31.1, T32.1, T46.5, Me1, Me2, Me14, Me15, Me16, Me17 and Me28) and eastern grey kangaroo (Macropus giganteus) (G16.1, G20.2, G26.4, G31.1) [41 (link)–43 (link)], as previously described [44 (link)]. The genotypes for 252 individual N. eugenii are given in S1 Table. Males were also genotyped at four tammar wallaby derived Y-linked microsatellite loci (MeY01, MeY28, MeY37A, MeY37B) [45 ] as previously described [22 ]. PCR products were analysed on a AB 3730 DNA Analyser (Applied Biosystems, USA) at the Ramaciotti Centre, UNSW (9 loci) or at AGRF Melbourne (7 loci), with the resultant DNA fragments sized using GeneMapper v3.7 (Applied Biosystems, USA).