A Reliable Molecular Method of Gender Determination for Mammals

Genetic differentiation of eastern wolves in Algonquin Park despite bridging gene flow between coyotes and grey wolves

“…Gender was determined either by amplification at the Zfx/Zfy intron (Shaw et al, 2003) or with Zfx/Sry primer pairs (Aasen and Medrano, 1990;Fain and LeMay, 1995). Confirmed males were then profiled at 4 Y chromosome microsatellite loci (MS34A, MS34B, MS41A and MS41B; Sundqvist et al, 2001) and at a 658 bp fragment of the Zfy intron with primers LGL-331 (5 0 -CAAATCATGCAAGGATAGAC-3 0 ; Shaw et al, 2003) and Yint2-335 (5 0 -GTCCATTGGATAATTCTTTCC-3 0 ; Shami, 2002).…”

“…Gender was determined either by amplification at the Zfx/Zfy intron (Shaw et al, 2003) or with Zfx/Sry primer pairs (Aasen and Medrano, 1990;Fain and LeMay, 1995). Confirmed males were then profiled at 4 Y chromosome microsatellite loci (MS34A, MS34B, MS41A and MS41B; Sundqvist et al, 2001) and at a 658 bp fragment of the Zfy intron with primers LGL-331 (5 0 -CAAATCATGCAAGGATAGAC-3 0 ; Shaw et al, 2003) and Yint2-335 (5 0 -GTCCATTGGATAATTCTTTCC-3 0 ; Shami, 2002). The polymerase chain reaction (PCR) chemical and cycling conditions for the Y chromosome microsatellite loci were as follows: For MS34, 5-10 ng of DNA was amplified in a 15 ml reaction with 1 Â PCR buffer, 0.2 mM dNTPs (Invitrogen, Burlington, Ontario, Canada), 1.5 mM MgCl 2 , 0.1 mM MS34A-F primer, 0.15 mM MS34B-F primer, 0.2 mM MS34-R primer and 1 U Taq DNA polymerase (Invitrogen).…”

Foraging ecology of ringed seals (Pusa hispida), beluga whales (Delphinapterus leucas) and narwhals (Monodon monoceros) in the Canadian High Arctic determined by stomach content and stable isotope analysis

“…The age of narwhals could not be estimated, but the lengths of their tusks ( !1 m) indicated that all three were adult males (Finley & Gibb 1982). The sex of ringed seals and toothed whales was determined from a skin sample using polymerase chain reactionbased genetic analysis (Shaw et al 2003).…”

Effective population size of an offshore population of bottlenose dolphins, Tursiops truncatus, from the São Pedro and São Paulo Archipelago, Brazil

“…DNA extraction followed the phenol/chloroform method described by Sambrook et al (1989) and modified by Shaw et al (2003). The sex of each individual was identified by the amplification of introns from ZFX and ZFY genes, which are located on the X and Y mammalian chromosomes, respectively (Page et al, 1987), using primers ZFY1204 and ZFY0097 under PCR conditions described in Pasbøll et al (1992).…”

“…For this reason, gender identification through analyses of molecular markers is becoming a powerful tool in the study of wild populations of cetaceans (e.g. Shaw et al, 2003;Oliveira et al, 2009). …”