Reliable and Robust Molecular Sexing of the Hen Harrier (Circus cyaneus) Using PCR-RFLP Analysis of the CHD 1 Gene


  • Supported by the School of Forensic & Investigative Sciences, University of Central Lancashire, and the Faculty of Science, University of Amsterdam.

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Arati Iyengar, Ph.D.

Department of Forensic & Investigative Science

University of Central Lancashire

Preston PR1 2HE, UK



The hen harrier (Circus cyaneus) is a bird of prey that is persecuted in the United Kingdom, and there is a need for a DNA-based individual identification and sexing system for the use in forensic investigations. This study reports a new set of PCR primers for the chromo-helicase-DNA-binding protein 1 gene, which allows sexing using PCR-RFLP. Instead of exonic primers that amplify across a large intron, this set consists of a primer within the intron, enabling reduction in amplicon sizes from 356 to 212 bp and 565 to 219 bp in W and Z chromosomes. DNA degradation and dilution experiments demonstrate that this set is significantly more robust than one that amplifies across the intron, and sequencing of the intronic primer-binding region across several individuals shows that it is highly conserved. While our objective is to incorporate this primer set into an STR-based individualization kit, it may in the meantime prove useful in forensic or conservation studies.