title: SRY sequence of Dasyurus geoffroii: adult male ear biopsy
design: Whole genomic DNA was extracted from an ear biopsy sample with a standard salting out method (Sunnucks and Hales 1996). The PCR reactions were performed in a 10 uL reaction using 3.75 uL of QIAGEN Multiplex PCR master mix, two uL deionised water, one uL of tissue DNA (five ng/uL) and 0.75 uL of two uM primer mix. Reactions were run under the following conditions: 15 min at 95C, followed by 40 cycles of 30 sec at 94C, 90 sec at the annealing temperature of 60C and 60 sec at 72C, concluding with 30 min at 60C. The 25 uL PCR products of five males were sequenced in an ABI 3730 sequencer (Murdoch University, Western Australia), and edited using Geneious Prime version 2023.2.1 (Biomatters Ltd, Auckland, New Zealand).