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Reliable and Robust Molecular Sexing of the Hen Harrier ( Circus cyaneus ) Using PCR ‐ RFLP Analysis of the CHD 1 Gene
Author(s) -
Henderson Anique,
Lee Christine Michelle,
Mistry Vanisha,
Thomas Martin Derek,
Iyengar Arati
Publication year - 2013
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/1556-4029.12030
Subject(s) - sexing , amplicon , primer (cosmetics) , biology , genetics , intron , polymerase chain reaction , gene , computational biology , microbiology and biotechnology , chemistry , organic chemistry
The hen harrier ( C ircus 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.