Molecular sexing
We determined the sex of individuals using a polymerase chain reaction (PCR) protocol adapted from Fridolfsson and Ellegren (1999). We used 0.25μL forward primer 2550F, 0.25μL reverse primer 2718Rand 6.25μL OneTaq® DNA Polymerase (New England BioLabs , Victoria, Australia) in combination with 1μL DNA for each of the 12.5μL reactions. We used an Eppendorf® Mastercycler machine with an annealing temperature of 48°C. To visualise the reactions, we ran the PCRs at 100V for 30 minutes on a 1.5% agarose gel with sybrstain (Invitrogen , NSW, Australia). Males were identified through the amplification of a single product while two products were visible for females.